Patent Publication Number: US-2022227825-A1

Title: Fgf21 variants

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
     This application is a continuation of U.S. patent application Ser. No. 15/776,726, filed May 16, 2018, which is a 35 U.S.C. § 371 filing of International Patent Application No. PCT/EP2016/079551, filed Dec. 2, 2016, which claims priority to European Patent Application No. 15306913.3, filed Dec. 2, 2015, the entire contents of which are incorporated herein by reference. 
    
    
     TECHNICAL FIELD OF THE INVENTION 
     The present invention relates to polypeptide variants of human fibroblast growth factor 21 (FGF21) and fusion molecules thereof, as well as to nucleic acid molecules encoding the same. It further relates to their use as medicaments, in particular for the treatment of obesity, overweight, metabolic syndrome, diabetes mellitus, hyperglycemia, dyslipidemia, non-alcoholic steatohepatitis (NASH) and/or atherosclerosis. 
     BACKGROUND OF THE INVENTION 
     Fibroblast growth factors (FGFs) are polypeptides widely expressed in developing and adult tissues. The FGF family currently consists of twenty-two members, FGF1 to FGF23 (one of them being referred to as FGF15/19). The members of the FGF family are highly conserved in both gene structure and amino acid sequence between vertebrate species.-FGF23) which are grouped into 6 subfamilies based on differences in sequence homology and phylogeny. The numbered ‘FGFs’ that are unassigned to subfamilies—the FGF homologous factors (previously known as FGF11-FGF14)—have high sequence identity with the FGF family but do not activate FGF receptors (FGFRs) and are therefore not generally considered members of the FGF family. While most of FGFs act as local regulators of cell growth and differentiation, recent studies indicated that FGF19 subfamily members including FGF15/19, FGF21 and FGF23 exert important metabolic effects by an endocrine fashion. The members of the FGF19 subfamily regulate diverse physiological processes that are not affected by classical FGFs. The wide variety of metabolic activities of these endocrine factors include the regulation of the bile acid, carbohydrate and lipid metabolism as well as phosphate, calcium and vitamin D homeostasis. 
     FGF21 was originally isolated from mouse embryos. FGF21 mRNA was most abundantly expressed in the liver, and to lesser extent in the thymus. Human FGF21 is highly similar (approximately 81% amino acid identity) to mouse FGF21. Among human FGF family members, FGF21 is the most similar (approximately 30% amino acid identity) to FGF19. FGF21 does not exhibit the proliferative and tumorigenic effectsthat are typical for a majority of the members of the FGF family. 
     FGF21 is a metabolic regulator produced primarily by the liver that exerts potent antidiabetic and lipid-lowering effects in animal models of obesity and type 2 diabetes mellitus. This hormone contributes to body weight regulation and is involved in the response to nutritional deprivation and ketogenic state in mice. The principal sites of metabolic actions of FGF21 are adipose tissue, liver and pancreas. Experimental studies have shown improvements in diabetes compensation and dyslipidemia after FGF21 administration in diabetic mice and primates. FGF21 has been shown to stimulate glucose uptake in mouse 3T3-L1 adipocytes in the presence and absence of insulin, and to decrease fed and fasting blood glucose, triglycerides, and glucagon levels in ob/ob and db/db mice and 8 week old ZDF rats in a dose dependant manner, thus, providing the basis for the use of FGF21 as a therapy for treating diabetes and obesity). 
     The administration of FGF21 to obese leptin-deficient ob/ob and leptin receptor-deficient db/db mice and obese ZDF rats significantly lowered blood glucose and triglycerides, decreased fasting insulin levels and improved glucose clearance during an oral glucose tolerance test. FGF21 did not affect food intake or body weight/composition of diabetic or lean mice and rats over the course of 2 weeks of administration. Importantly, FGF21 did not induce mitogenicity, hypoglycemia, or weight gain at any dose tested in diabetic or healthy animals or when overexpressed in transgenic mice. FGF21-overexpressing transgenic mice were resistant to diet-induced obesity. 
     The administration of FGF21 to diabetic rhesus monkeys for 6 weeks reduced fasting plasma glucose, fructosamine, triglyceride, insulin and glucagon levels. Importantly, hypoglycemia was not observed during the study despite of significant glucose-lowering effects. FGF21 administration also significantly lowered LDL-cholesterol and increased HDL-cholesterol and, in contrast to mice, slightly but significantly decreased body weight. 
     SUMMARY OF THE INVENTION 
     A problem associated with the use of human wild-type FGF21 as a therapeutic in the treatment of diabetes, obesity, metabolic syndrome and other indications is its limited half-life in vivo. In mice FGF21 has a half-life below 1 hour and in cynomolgus monkeys ( Macaca fascicularis ) around 2-4 hours only. Therefore, in developing an FGF21 protein for use as a therapeutic there is a need for variants with improved pharmaceutical properties over human wild-type FGF21, in particular increased stability against proteases and/or thermal degradation and/or increased potency/efficacy. To achieve this goal, FGF21 variants were designed to identify FGF21 polypeptides having enhanced stability. 
     In a first aspect, the present invention relates to a variant of human fibroblast growth factor 21 (FGF21) comprising the amino acid sequence of SEQ ID NO: 171, 
     wherein
         amino acids 1-28 are deleted;   Xaa55 is Q or another amino acid, such as G or C;   Xaa147 is P or C, or deleted;   Xaa148 is G or C, or deleted;   Xaa149 is N or C, or deleted;   Xaa150 is K or another amino acid, such as H, or deleted;   Xaa151 is S or deleted;   Xaa152 is P or another amino acid, such as A or L, or deleted;   Xaa153 is H or another amino acid, such as Q, Y or K, or deleted;   Xaa154 is R or another amino acid, such as Q or K, or deleted;   Xaa155 is D or another amino acid, such as L, K, Y, P, E, N or C, or deleted;   Xaa156 is P or another amino acid, such as A or K, or deleted;   Xaa157 is A or another amino acid, such as V, G or S, or deleted;   Xaa158 is P or another amino acid, such as H, or deleted;   Xaa159 is R or another amino acid, such as H, K or Q, or deleted;   Xaa160 is G or deleted;   Xaa161 is P or C, or deleted;   Xaa162 is A or another amino acid, such as C or Y, or deleted;   Xaa163 is R or another amino acid, such as H, or deleted;   Xaa184 is Q or deleted;   Xaa185 is P or deleted;   Xaa186 is P or deleted;   Xaa187 is D or deleted;   Xaa188 is V or deleted;   Xaa189 is G or deleted;   Xaa190 is S or deleted;   Xaa191 is S or deleted;   Xaa192 is D or deleted;   Xaa193 is P or deleted;   Xaa194 is L or deleted;   Xaa195 is S or C, or deleted;   Xaa196 is M or another amino acid, such as P, V or C, or deleted;   Xaa197 is V or another amino acid, such as E, D, G or M, or deleted;   Xaa198 is G or another amino acid, such as E, D, R, K, Y, P or V, or deleted;   Xaa199 is P or another amino acid, such as S, Q, R, T, G, F, L, D or M, or deleted;   Xaa200 is S or another amino acid, such as Q, M, C, P, N or H, or deleted;   Xaa201 is Q or another amino acid, such as P or S, or deleted;   Xaa202 is G or another amino acid, such as T, or deleted;   Xaa203 is R or another amino acid, such as E, H or C, or deleted;   Xaa204 is S or deleted;   Xaa205 is P or deleted;   Xaa206 is S or deleted; and   Xaa207 is Y or deleted;
 
wherein, optionally, SEQ ID NO: 171 comprises a substitution of at least one of the following amino acids with C: R47, L49, T51, A54, Q56, A59, H60, E62, I63, G67, V69, G71, A72, A73, S76, P77, E78, S79, L80, L81, Q82, L83, I91, L94, G95, V96, K97, T98, R100, L102, Q104, D107, G108, L110, G112, L114, A120, R124, D130, Y132, Q136, S137, A139, H140, L142, P143, H145, L146, L165, L167, L170, P174;
 
with the proviso that the variant of human FGF21 is not mature human wild-type FGF21 (SEQ ID NO: 2) and comprises 0, 2, 4, 6 or 8 additional cysteines as compared to mature human wild-type FGF21 (SEQ ID NO: 2),
 
wherein, optionally, SEQ ID NO: 171 further comprises the mutation G141S and/or the mutation P174L.
       

     In one embodiment,
         Xaa147 to Xaa159 are deleted and replaced by a protease resistant peptide linker, such as the peptide linker of SEQ ID NO: 161 or SEQ ID NO: 162;   Xaa149 to Xaa155 are deleted and replaced by a protease resistant peptide linker, such as the peptide linker of SEQ ID NO: 161, SEQ ID NO: 163 or SEQ ID NO: 164;   Xaa149 to Xaa159 are deleted and replaced by a protease resistant peptide linker, such as the peptide linker of SEQ ID NO: 161 or SEQ ID NO: 162;   Xaa149 to Xaa162 are deleted and replaced by a protease resistant peptide linker, such as the peptide linker of SEQ ID NO: 163 or SEQ ID NO: 165;   Xaa149 to Xaa163 are deleted and replaced by a protease resistant peptide linker, such as the peptide linker of SEQ ID NO: 161;   Xaa159 to Xaa163 are deleted and replaced by a protease resistant peptide linker, such as GA, GY, HH, GE or HE;   Xaa197 to Xaa203 are deleted and replaced by a protease resistant peptide linker, such as the peptide linker of SEQ ID NO: 166 or SEQ ID NO: 167;   a protease resistant peptide linker, such as the peptide linker of SEQ ID NO: 168, is inserted between Xaa198 and Xaa199; and/or   the amino acid sequence of SEQ ID NO: 169 or SEQ ID NO: 170 is added after S209 of SEQ ID NO: 171.       

     In one embodiment, the varant of human FGF21 has at least 90% or at least 91% or at least 92% or at least 93% or at least 94% or at least 95% or at least 96% or at least 97% or at least 98% sequence identity with mature human wild-type FGF21 (SEQ ID NO: 2). 
     In one embodiment,
         Xaa55 is Q or C;   Xaa147 is P or C;   Xaa148 is G or C;   Xaa149 is N or C;   Xaa150 is K;   Xaa151 is S;   Xaa152 is P;   Xaa153 is H;   Xaa154 is R;   Xaa155 is D or C;   Xaa156 is P;   Xaa157 is A;   Xaa158 is P;   Xaa159 is R;   Xaa160 is G;   Xaa161 is P or C;   Xaa162 is A or C;   Xaa163 is R;   Xaa184 is Q;   Xaa185 is P;   Xaa186 is P;   Xaa187 is D;   Xaa188 is V;   Xaa189 is G;   Xaa190 is S;   Xaa191 is S;   Xaa192 is D;   Xaa193 is P;   Xaa194 is L;   Xaa195 is S or C;   Xaa196 is M or another amino acid, such as P, V or C, or deleted;   Xaa197 is V or another amino acid, such as E, D, G or M, or deleted;   Xaa198 is G or another amino acid, such as E, D, R, K, Y, P or V, or deleted;   Xaa199 is P or another amino acid, such as S, Q, R, T, G, F, L, D or M, or deleted;   Xaa200 is S or another amino acid, such as Q, M, C, P, N or H, or deleted;   Xaa201 is Q or another amino acid, such as P or S, or deleted;   Xaa202 is G or another amino acid, such as T, or deleted;   Xaa203 is R or another amino acid, such as E, H or C, or deleted;   Xaa204 is S;   Xaa205 is P;   Xaa206 is S; and   Xaa207 is Y;
 
wherein, optionally, SEQ ID NO: 171 comprises a substitution of at least one of the following amino acids with C: R47, L49, T51, A54, Q56, A59, H60, E62, 163, G67, V69, G71, A72, A73, S76, P77, E78, S79, L80, L81, Q82, L83, I91, L94, G95, V96, K97, T98, R100, L102, Q104, D107, G108, L110, G112, L114, A120, R124, D130, Y132, Q136, S137, A139, H140, L142, P143, H145, L146, L165, L167, L170, P174.
       

     In one embodiment, the variant comprises the amino acid sequence of SEQ ID NO: 175 (wherein the variables are as defined herein), 
     wherein, optionally, SEQ ID NO: 175 comprises a substitution of at least one of the following amino acids with C: R47, L49, T51, A54, Q56, A59, H60, E62, 163, G67, V69, G71, A72, A73, S76, P77, E78, S79, L80, L81, Q82, L83, I91, L94, G95, V96, K97, T98, R100, L102, Q104, D107, G108, L110, G112, L114, A120, R124, D130, Y132, Q136, S137, A139, H140, L142, P143, H145, L146, L165, L167, L170, P174;
 
with the proviso that SEQ ID NO: 175 is not mature human wild-type FGF21 (SEQ ID NO: 2) and comprises 0, 2, 4, 6 or 8 additional cysteines as compared to mature human wild-type FGF21 (SEQ ID NO: 2),
 
wherein, optionally, SEQ ID NO: 175 further comprises the mutation G141S and/or the mutation P174L.
 
     In one embodiment,
         Xaa55 is Q or C;   Xaa147 is P or C;   Xaa148 is G;   Xaa149 is N or C;   Xaa155 is D or C;   Xaa161 is P;   Xaa162 is A;   Xaa195 is S;   Xaa196 is M;   Xaa197 is V;   Xaa198 is G or another amino acid, such as E, D, R, K, Y, P or V, or deleted;   Xaa199 is P or another amino acid, such as S, Q, R, T, G, F, L, D or M, or deleted;   Xaa200 is S;   Xaa201 is Q;   Xaa202 is G; and   Xaa203 is R;
 
wherein, optionally, SEQ ID NO: 171 comprises a substitution of at least one of the following amino acids with C: R47, A59, H60, G71, S76, S79, D107, G108, L142, P174.
       

     In one embodiment, the variant comprises the amino acid sequence of SEQ ID NO: 176 (wherein the variables are as defined herein), 
     wherein, optionally, SEQ ID NO: 176 comprises a substitution of at least one of the following amino acids with C: R47, A59, H60, G71, S76, S79, D107, G108, L142, P174;
 
with the proviso that SEQ ID NO: 176 is not mature human wild-type FGF21 (SEQ ID NO: 2) and comprises 0, 2, 4, 6 or 8 additional cysteines as compared to mature human wild-type FGF21 (SEQ ID NO: 2),
 
wherein, optionally, SEQ ID NO: 176 further comprises the mutation G141S and/or the mutation P174L.
 
     In one embodiment, Xaa198 is G, R, K or Y. 
     In one embodiment, Xaa199 is P or deleted. 
     In one embodiment, SEQ ID NO: 171, 175 or 176 comprises a substitution of the following amino acids with C:
         R47 and P174;   Xaa55 and Xaa147;   Xaa55 and Xaa149;   A59 and G71;   H60 and S79;   S76 and S79;   D107 and Xaa155; and/or   G108 and L142.       

     In one embodiment, SEQ ID NO: 171, 175 or 176 comprises a substitution of the following amino acids with C:
         Xaa55 and Xaa147;   Xaa55 and Xaa149;   A59 and G71;   S76 and S79;   D107 and Xaa155; or   G108 and L142.       

     In one embodiment, SEQ ID NO: 171, 175 or 176 comprises a substitution of Xaa55 and Xaa147 with C. In another embodiment, SEQ ID NO: 171, 175 or 176 comprises a substitution of Xaa55 and Xaa149 with C. 
     In one embodiment,
         Xaa55 is C, Xaa147 is C, Xaa149 is N, Xaa155 is D, Xaa198 is G, and Xaa199 is deleted;   Xaa55 is C, Xaa147 is C, Xaa149 is N, Xaa155 is D, Xaa198 is Y, and Xaa199 is P;   Xaa55 is C, Xaa147 is P, Xaa149 is C, Xaa155 is D, Xaa198 is G, and Xaa199 is deleted;   Xaa55 is C, Xaa147 is P, Xaa149 is C, Xaa155 is D, Xaa198 is Y, and Xaa199 is P;   Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is D, Xaa198 is G, and Xaa199 is deleted;   Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is D, Xaa198 is R, and Xaa199 is P;   Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is D, Xaa198 is K, and Xaa199 is P;   Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is D, Xaa198 is Y, and Xaa199 is P;   Xaa55 is C, Xaa147 is C, Xaa149 is N, Xaa155 is D, Xaa198 is G, and Xaa199 is P;   Xaa55 is C, Xaa147 is P, Xaa149 is C, Xaa155 is D, Xaa198 is G, and Xaa199 is P;   Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is D, Xaa198 is G, and Xaa199 is deleted, wherein SEQ ID NO: 171, 175 or 176 comprises the substitution of A59 and G71 with C;   Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is D, Xaa198 is Y, and Xaa199 is P, wherein SEQ ID NO: 171, 175 or 176 comprises the substitution of A59 and G71 with C;   Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is D, Xaa198 is G, and Xaa199 is deleted, wherein SEQ ID NO: 171, 175 or 176 comprises the substitution of S76 and S79 with C;   Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is D, Xaa198 is Y, and Xaa199 is P, wherein SEQ ID NO: 171, 175 or 176 comprises the substitution of S76 and S79 with C;   Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is D, Xaa198 is G, and Xaa199 is deleted, wherein SEQ ID NO: 171, 175 or 176 comprises the substitution of G108 and L142 with C;   Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is D, Xaa198 is Y, and Xaa199 is P, wherein SEQ ID NO: 171, 175 or 176 comprises the substitution of G108 and L142 with C;   Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is C, Xaa198 is G, and Xaa199 is deleted, wherein SEQ ID NO: 171, 175 or 176 comprises the substitution of D107 with C; or   Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is C, Xaa198 is Y, and Xaa199 is P, wherein SEQ ID NO: 171, 175 or 176 comprises the substitution of D107 with C.       

     In one embodiment, the variant further comprises at least one additional amino acid at its N-terminus. 
     In one embodiment, the at least one additional amino acid is selected from the group consisting of G, A, N and C. 
     In a second aspect, the present invention relates to a variant of human FGF21 comprising or consisting of an amino acid sequence according to one of SEQ ID NOs: 1 to 3, 172, 173 and 174, 
     wherein the amino acid sequence comprises at least one of the following mutations: 
     Q55G, K150H, P152A, P152L, P152 deleted, H153Q, H153Y, H153K, H153 deleted, R154Q, R154K, D155L, D155K, D155Y, D155P, D155E, D155N, P156A, P156K, A157V, A157G, A157S, P158H, R159H, R159K, R159Q, A162Y, R163H, Q184 deleted, P185 deleted, P186 deleted, D187 deleted, V188 deleted, G189 deleted, S190 deleted, S191 deleted, D192 deleted, P193 deleted, L194 deleted, S195 deleted, M196 deleted, M196P, M196V, V197E, V197D, V197 deleted, V197G, V197M, G198E, G198D, G198R, G198K, G198Y, G198P, G198V, G198 deleted, P199S, P199Q, P199 deleted, P199R, P199T, P199G, P199F, P199L, P199D, P199M, S200Q, S200M, S200P, S200N, S200H, S200 deleted, Q201P, Q201S, Q201 deleted, G202T, G202 deleted, R203E, R203H, R203 deleted, S204 deleted, P205 deleted, S206 deleted, Y207 deleted, P147-R159 replaced by GSGS (SEQ ID NO: 161), P147-R159 replaced by GGSGGS (SEQ ID NO: 162), N149-D155 replaced by GSGS (SEQ ID NO: 161), N149-D155 replaced by GSHSG (SEQ ID NO: 163), N149-D155 replaced by ATTS (SEQ ID NO: 164), N149-R159 replaced by GSGS (SEQ ID NO: 161), N149-R159 replaced by GGSGGS (SEQ ID NO: 162), N149-A162 replaced by GSHSG (SEQ ID NO: 163), N149-A162 replaced by GSHSGS (SEQ ID NO: 165), N149-R163 replaced by GSGS (SEQ ID NO: 161), R159-R163 replaced by GA, GY, HH, GE or HE, V197-R203 replaced by GHRSHLQTVF (SEQ ID NO: 166), V197-R203 replaced by GLNSMV (SEQ ID NO: 167), GGGGS (SEQ ID NO: 168) inserted between G198 and P199, PLSMVGPSQGRSPSYAS (SEQ ID NO: 169) or PLSMVGSQGRSPSYAS (SEQ ID NO: 170) added after S209; 
     and/or
 
wherein the amino acid sequence comprises a substitution of two, four, six or eight of the following amino acids with C:
 
     R47, L49, T51, A54, Q55, Q56, A59, H60, E62, I63, G67, V69, G71, A72, A73, S76, P77, E78, S79, L80, L81, Q82, L83, I91, L94, G95, V96, K97, T98, R100, L102, Q104, D107, G108, L110, G112, L114, A120, R124, D130, Y132, Q136, S137, A139, H140, L142, P143, H145, L146, P147, G148, N149, D155, P161, A162, L165, L167, L170, P174, S195, M196, S200, R203; 
     wherein the numbering of the amino acids is in accordance with SEQ ID NO: 1. 
     In one embodiment, the amino acid sequence comprises at least one of the following mutations: 
     K150H, P152A, P152L, P152 deleted, H153Q, H153Y, H153K, H153 deleted, R154Q, R154K, D155L, D155K, D155Y, D155P, D155E, D155N, P156A, P156K, A157V, A157G, A157S, P158H, R159H, R159K, R159Q, A162Y, R163H, M196 deleted, M196P, M196V, V197E, V197D, V197 deleted, V197G, V197M, G198E, G198D, G198R, G198K, G198Y, G198P, 
     G198V, G198 deleted, P199S, P199Q, P199 deleted, P199R, P199T, P199G, P199F, P199L, P199D, P199M, S200Q, S200M, S200P, S200N, S200H, S200 deleted, Q201P, Q201S, Q201 deleted, G202T, G202 deleted, R203E, R203H, R203 deleted; 
     and/or
 
the amino acid sequence comprises a substitution of two, four, six or eight of the following amino acids with C:
 
     R47, L49, T51, A54, Q55, Q56, A59, H60, E62, I63, G67, V69, G71, A72, A73, S76, P77, E78, S79, L80, L81, Q82, L83, I91, L94, G95, V96, K97, T98, R100, L102, Q104, D107, G108, L110, G112, L114, A120, R124, D130, Y132, Q136, S137, A139, H140, L142, P143, H145, L146, P147, G148, N149, D155, P161, A162, L165, L167, L170, P174, S195, M196, S200, R203. 
     In one embodiment, the amino acid sequence comprises at least one of the following mutations: M196 deleted, M196P, M196V, V197E, V197D, V197 deleted, V197G, V197M, G198E, G198D, G198R, G198K, G198Y, G198P, G198V, G198 deleted, P199S, P199Q, P199 deleted, P199R, P199T, P199G, P199F, P199L, P199D, P199M, S200Q, S200M, S200P, S200N, S200H, S200 deleted, Q201P, Q201S, Q201 deleted, G202T, G202 deleted, R203E, R203H, R203 deleted; 
     and/or
 
the amino acid sequence comprises a substitution of two, four, six or eight of the following amino acids with C:
 
     R47, L49, T51, A54, Q55, Q56, A59, H60, E62, 163, G67, V69, G71, A72, A73, S76, P77, E78, S79, L80, L81, Q82, L83, 191, L94, G95, V96, K97, T98, R100, L102, Q104, D107, G108, L110, G112, L114, A120, R124, D130, Y132, Q136, S137, A139, H140, L142, P143, H145, L146, P147, G148, N149, D155, P161, A162, L165, L167, L170, P174. 
     In one embodiment, the amino acid sequence comprises a substitution of the following amino acids with C:
         R47 and P174;   Q55 and P147;   Q55 and N149;   A59 and G71;   H60 and S79;   S76 and S79;   D107 and D155; and/or   G108 and L142.       

     In one embodiment, the amino acid sequence comprises a mutation selected from the group consisting of G198R, G198K, G198Y and P199 deleted; 
     and/or
 
the amino acid sequence comprises a substitution of the following amino acids with C:
         Q55 and P147;   Q55 and N149;   A59 and G71;   S76 and S79;   D107 and D155; or   G108 and L142.       

     In one embodiment, the amino acid sequence comprises a substitution of Q55 and P147 with C. 
     In another embodiment, the amino acid sequence comprises a substitution of Q55 and N149 with C. 
     In one embodiment, the amino acid sequence comprises at least one further mutation (e.g., 1, 2, 3, 4 or 5 further mutation(s)) at another site of one of SEQ ID NOs: 1 to 3, 172, 173 and 174. 
     In one embodiment, the amino acid sequence has at least 90% or at least 91% or at least 92% or at least 93% or at least 94% or at least 95% or at least 96% or at least 97% or at least 98% sequence identity with one of SEQ ID NOs: 1 to 3, 172, 173 and 174. 
     In a third aspect, the present invention relates to a variant of human FGF21 comprising or consisting of an amino acid sequence according to one of SEQ ID NOs: 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327 and 328, wherein, optionally, the variant further comprises the mutation G141S and/or the mutation P174L, wherein the numbering of the amino acids is in accordance with SEQ ID NO: 1. 
     In one embodiment, the variant according to the first, second and third aspect has an increased proteolytic stability in human and/or murine blood plasma as compared to mature human wild-type FGF21 (SEQ ID NO: 2). 
     In one embodiment, the variant has an increased thermal stability as compared to mature human wild-type FGF21 (SEQ ID NO: 2). 
     In one embodiment, the variant induces phosphorylation of the mitogen-activated protein kinase (MAPK) ERK1/2. 
     In one embodiment, the variant induces phosphorylation of the mitogen-activated protein kinase (MAPK) ERK1/2 with an EC50 of 100 nmol/L or lower, or 90 nmol/L or lower, or 80 nmol/L or lower, or 70 nmol/L or lower, or 60 nmol/L or lower, or 50 nmol/L or lower, or 40 nmol/L or lower, or 30 nmol/L or lower, or 20 nmol/L or lower, or 15 nmol/L or lower, or 12 nmol/L or lower, or 11 nmol/L or lower, or 10 nmol/L or lower, or 9 nmol/L or lower, or 8 nmol/L or lower, or 7 nmol/L or lower, or 6 nmol/L or lower, or 5 nmol/L or lower, or 4 nmol/L or lower, or 3 nmol/L or lower, or 2 nmol/L or lower, e.g., as determined in an In-Cell Western (ICW) assay, such as in a mammalian cell culture. 
     In one embodiment, the variant further comprises at least one label or tag allowing the detection and/or isolation of the variant. 
     In one embodiment, the variant is fused or conjugated to a half-life extension module. 
     In one embodiment, the half-life extension module is selected from the group consisting of a polymer (e.g., polyethylene glycol (PEG), hydroxyethyl starch (HES), hyaluronic acid, polysialic acid), an unstructured (poly-)peptide chain (e.g., PAS, XTEN), an elastin-like polypeptide (ELP), a serum protein (e.g., albumin), a serum protein binding molecule (e.g., an albumin binding domain (ABD), an albumin binding fatty acid), an antibody, an immunoglobulin, an Fc region/domain of an immunoglobulin and an immunoglobulin binding domain. 
     In another aspect, the present invention relates to a fusion molecule comprising a variant of human FGF21 as defined above and at least one other active pharmaceutical ingredient. 
     In another aspect, the present invention relates to a nucleic acid molecule encoding a variant of human FGF21 as defined above or a fusion molecule as defined above. 
     In one embodiment, the nucleic acid molecule is contained in a vector or is integrated into a genome. 
     In another aspect, the present invention relates to a host cell containing a nucleic acid molecule as defined above. 
     In another aspect, the present invention relates to a method of producing a variant of human FGF21 as defined above or a fusion molecule as defined above, comprising cultivating a host cell as defined above and isolating the variant or fusion molecule from the culture medium. 
     In another aspect, the present invention relates to a pharmaceutical composition comprising a variant of human FGF21 as defined above or a fusion molecule as defined above or a nucleic acid molecule as defined above or a host cell as defined above, together with a pharmaceutically acceptable carrier and/or excipient. 
     In one embodiment, the pharmaceutical composition further comprises at least one other active pharmaceutical ingredient. 
     In another aspect, the present invention relates to a combination of a variant of human FGF21 as defined above with at least one other active pharmaceutical ingredient. 
     In one embodiment, the at least one other active pharmaceutical ingredient comprised in the fusion molecule or the pharmaceutical composition or the combination as defined above is selected from the group consisting of insulin and insulin derivatives, GLP-1, GLP-1 analogues and GLP-1 receptor agonists, polymer bound GLP-1 and GLP-1 analogues, dual GLP-1/GIP agonists, dual GLP-1/glucagon receptor agonists, PYY3-36 or analogues thereof, pancreatic polypeptide or analogues thereof, glucagon receptor agonists or antagonists, GIP receptor agonists or antagonists, ghrelin antagonists or inverse agonists, xenin and analogues thereof, peptides (specifically) binding to one of the aforementioned receptors (e.g., GLP-1 receptor, GIP receptor, glucagon receptor), DDP-IV inhibitors, SGLT-2 inhibitors, dual SGLT-2/SGLT-1 inhibitors, biguanides, thiazolidinediones, PPAR agonists, PPAR modulators, sulfonylureas, meglitinides, alpha-glucosidase inhibitors, amylin and amylin analogues, GPR119 agonists, GPR40 agonists, GPR120 agonists, GPR142 agonists, TGR5 agonists, AMPK stimulants, AMPK activators, inhibitors of 11-beta-HSD, activators of glucokinase, inhibitors of DGAT, inhibitors of protein tyrosine phosphatase 1, inhibitors of glucose-6-phosphatase, inhibitors of fructose-1,6-bisphosphatase, inhibitors of glycogen phosphorylase, inhibitors of phosphoenol pyruvate carboxykinase, inhibitors of glycogen synthase kinase, inhibitors of pyruvate dehydrogenase kinase, CCR-2 antagonists, modulators of glucose transporter-4, somatostatin receptor 3 agonists, HMG-CoA-reductase inhibitors, fibrates, nicotinic acid and derivatives thereof, nicotinic acid receptor 1 agonists, ACAT inhibitors, cholesterol absorption inhibitors, bile acid-binding substances, IBAT inhibitors, MTP inhibitors, modulators of PCSK9, LDL receptor up-regulators (liver selective thyroid hormone receptor beta agonists), HDL-raising compounds, lipid metabolism modulators, PLA2 inhibitors, ApoA-I enhancers, cholesterol synthesis inhibitors, omega-3 fatty acids and derivatives thereof, active substances for the treatment of obesity, CB1 receptor antagonists, MCH-1 antagonists, MC4 receptor agonists and partial agonists, NPY5 or NPY2 antagonists, NPY4 agonists, beta-3 adrenergic receptor agonists, leptin or leptin mimetics, 5HT2c receptor agonists, lipase inhibitors, angiogenesis inhibitors, H3 antagonists, AgRP inhibitors, triple monoamine uptake inhibitors, MetAP2 inhibitors, antisense oligonucleotides against production of fibroblast growth factor receptor 4 or prohibitin targeting peptide-1, drugs for influencing high blood pressure, chronic heart failure or atherosclerosis, angiotensin II receptor antagonists, dual angiotensin receptor blockers (ARB), angiotensin converting enzyme (ACE) inhibitors, angiotensin converting enzyme 2 (ACE-2) activators, renin inhibitors, prorenin inhibitors, endothelin converting enzyme (ECE) inhibitors, endothelin receptor blockers, endothelin antagonists, diuretics, aldosterone antagonists, aldosterone synthase inhibitors, alpha-blockers, antagonists of the alpha-2 adrenergic receptor, beta-blockers, mixed alpha-/beta-blockers, calcium antagonists/calcium channel blockers (CBBs), dual mineralocorticoid/CCBs, centrally acting antihypertensives, inhibitors of neutral endopeptidase, aminopeptidase-A inhibitors, vasopeptide inhibitors, dual vasopeptide inhibitors, neprilysin-ACE inhibitors, neprilysin-ECE inhibitors, dual-acting Angiotensin (AT) receptor-neprilysin inhibitors, dual AT1/endothelin-1 (ETA) antagonists, advanced glycation end-product breakers, recombinant renalase, blood pressure vaccines, anti-RAAS vaccines, AT1- or AT2-vaccines, modulators of genetic polymorphisms with antihypertensive response and thrombocyte aggregation inhibitors. 
     In one embodiment, the at least one other active pharmaceutical ingredient is a glucagon-like peptide-1 (GLP-1) receptor agonist. 
     In another aspect, the present invention relates to a kit comprising a variant of human FGF21 as defined above or a fusion molecule as defined above or a nucleic acid molecule as defined above or a host cell as defined above or a pharmaceutical composition as defined above. 
     In another aspect, the present invention relates to a variant of human FGF21 as defined above or a fusion molecule as defined above or a nucleic acid molecule as defined above or a host cell as defined above or a pharmaceutical composition as defined above for use as a medicament. 
     In another aspect, the present invention relates to a variant of human FGF21 as defined above or a fusion molecule as defined above or a nucleic acid molecule as defined above or a host cell as defined above or a pharmaceutical composition as defined above for use in the treatment of a disease or disorder selected from the group consisting of obesity, overweight, metabolic syndrome, diabetes mellitus, hyperglycemia, dyslipidemia, non-alcoholic steatohepatitis (NASH) and atherosclerosis. 
     In another aspect, the present invention relates to the use of a variant of human FGF21 as defined above or a fusion molecule as defined above or a nucleic acid molecule as defined above or a host cell as defined above or a pharmaceutical composition as defined above in the manufacture of a medicament for the treatment of a disease or disorder selected from the group consisting of obesity, metabolic syndrome, diabetes mellitus, hyperglycemia, dyslipidemia, non-alcoholic steatohepatitis (NASH) and atherosclerosis. 
     In another aspect, the present invention relates to a method of treating a disease or disorder selected from the group consisting of obesity, metabolic syndrome, diabetes mellitus, hyperglycemia, dyslipidemia, non-alcoholic steatohepatitis (NASH) and atherosclerosis, the method comprising administering a variant of human FGF21 as defined above or a fusion molecule as defined above or a nucleic acid molecule as defined above or a host cell as defined above or a pharmaceutical composition as defined above to a subject in need thereof. 
     In one embodiment, the disease or disorder is diabetes mellitus. 
    
    
     DESCRIPTION OF THE FIGURES 
       FIGS. 1A-1L  show mass spectra and deconvolution data for FGF21 of SEQ ID NO: 3 and its metabolites. Mass spectra ( FIG. 1A ) and deconvolution ( FIG. 1B ) of the sequence of amino acids [1-182] of SEQ ID NO: 3; Mass spectra ( FIG. 1C ) and deconvolution ( FIG. 1D ) of the sequence of amino acids [1-172] of SEQ ID NO: 3 in human plasma (also found in rat and murine plasma); RT: 33.6 min; the multicharged masses labelled with C were deconvoluted to 18,429 Da; Mass spectra ( FIG. 1E ) and deconvolution ( FIG. 1F ) of the sequence of amino acids [132-182] of SEQ ID NO: 3 in human plasma (also found in rat plasma); RT: 30.4 min; the multicharged masses labelled with F were deconvoluted to 5,113.67 Da; Mass spectra ( FIG. 1G ) and deconvolution ( FIG. 1H ) of the proposed sequences of amino acids [129-182], [125-177], [126-178] or [124-176] of SEQ ID NO: 3 in human plasma; RT: 30.8 min; the multicharged masses labelled with E were deconvoluted to 5,379 Da; Mass spectra ( FIG. 11 ) and deconvolution ( FIG. 1J ) of the proposed sequence of amino acids [136-182] of SEQ ID NO: 3 in human plasma; RT: 31.3 min; the multicharged masses labelled with B were deconvoluted to 4,732.41 Da; Mass spectra ( FIG. 1K ) and deconvolution ( FIG. 1L ) of the proposed sequence of amino acids [53-129] or [21-98] of SEQ ID NO: 3 in murine plasma; RT: 32.5; the multicharged masses labelled with A were deconvoluted to 8,564.67 Da. 
       FIG. 2  shows an analysis of the stability of mature human FGF21 (SEQ ID NO: 2) and FGF21 variants in human plasma. The proteins were incubated in plasma for 24 hours at 37° C., and remaining FGF21 amounts were measured using an ELISA recognizing intact mature FGF21. 
     Amounts at the start of the incubation were set as 100%. Data are mean+SD. Corresponding SEQ ID NOs are shown in Table 3. GGGGS (SEQ ID NO: 168); GHRSHLQTVF (SEQ ID NO: 166). 
       FIGS. 3A-3W  show dose-response curves of ERK1/2-phosphorylation in CHO cells overexpressing human FGFR1c and beta-klotho after stimulus with mature human FGF21 (SEQ ID NO: 2) or various FGF21 variants measured via In-Cell Western ( FIGS. 3A to 3W ). Corresponding SEQ ID NOs are shown in Table 4.  FIG. 3A : GHRSHLQTVF (SEQ ID NO: 166); GLNSMV (SEQ ID NO: 167).  FIG. 3J : GSGS (SEQ ID NO: 161).  FIG. 3O : PLSMVGPSQGRSPSYAS (SEQ ID NO: 169). 
       FIGS. 4A-4B  show dose-response curves of ERK1/2-phosphorylation in primary human subcutaneous adipocytes after stimulus with mature human FGF21 (SEQ ID NO: 2) or various FGF21 variants measured via In-Cell Western ( FIGS. 4A and 4B ). Corresponding SEQ ID NOs are shown in Table 5. 
       FIG. 5  shows an analysis of the thermal stability of human FGF21 (SEQ ID NO: 3) and FGF21 variants analyzed via differential scanning fluorimetry (DSF). Exemplary melting curves from the ThermoFluorTM experiment for human FGF21 (SEQ ID NO: 3) and selected FGF21 variants are shown. Relative fluorescence units normalized at 25° C. are plotted against temperature (° C.). Corresponding SEQ ID NOs are shown in Table 6. 
       FIGS. 6A-6C  shows structural details of computational and experimental models of human FGF21. ( FIG. 6A ) Superposition of computational models represented as backbone structures based on different experimental homologous structures of FGF19 and FGF23, white (BL 2P23): model created with BioLuminate based on the structure of FGF19 (PDB code 2P23); light gray (SM 1PWA): model created using the Swiss-Model server based on the structure of FGF19 (PDB code 1PWA); dark gray (SM 2P23): model created using the Swiss-Model server based on the structure of FGF19 (PDB code 2P23); black (SM 2P39): model created using the Swiss-Model server based on the structure of FGF23 (PDB code 2P39). Particularly the loop region around residues 150-158 (encircled) is significantly different in these models due to the structural differences of the underlying experimental data. ( FIG. 6B ) Superposition, shown as a cartoon, of molecule A and molecule B present in the X-ray crystal structure of FGF21 Q55C, N149C (SEQ ID NO: 158). The engineered disulfide bond between cysteines 149 and 55 that stabilize the conformation, thereby increasing the thermal stability, is highlighted. ( FIG. 6C ) 2mFo-dFc electron density around the engineered C55-C149 disulfide link in molecule B. The contour level is la. Between C149 and P156, the electron density is discontinuous, suggesting significant disorder. 
     DETAILED DESCRIPTION OF THE INVENTION 
     Although the present invention is described in detail below, it is to be understood that this invention is not limited to the particular methodologies, protocols and reagents described herein as these may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention which will be limited only by the appended claims. Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art. 
     In the following, certain elements of the present invention will be described. These elements may be listed with specific embodiments, however, it should be understood that they may be combined in any manner and in any number to create additional embodiments. The variously described examples and embodiments should not be construed to limit the present invention to only the explicitly described embodiments. This description should be understood to support and encompass embodiments which combine the explicitly described embodiments with any number of the disclosed elements. Furthermore, any permutations and combinations of all described elements in this application should be considered disclosed by the description of the present application unless the context indicates otherwise. 
     The terms used herein are defined generally as described in “A multilingual glossary of biotechnological terms: (IUPAC Recommendations)”, H.G.W. Leuenberger, B. Nagel, and H. Kölbl, Eds., Helvetica Chimica Acta, CH-4010 Basel, Switzerland, (1995). 
     The practice of the present invention will employ, unless otherwise indicated, conventional methods of chemistry, biochemistry, cell biology, immunology, and recombinant DNA techniques which are explained in the literature in the field (Sambrook, J. et al. (2001) Molecular Cloning: A Laboratory Manual, 3rd Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.). 
     Throughout this specification and the claims which follow, unless the context requires otherwise, the word “comprise”, and variations such as “comprises” and “comprising”, will be understood to imply the inclusion of a stated member, integer or step or group of members, integers or steps but not the exclusion of any other member, integer or step or group of members, integers or steps although in some embodiments such other member, integer or step or group of members, integers or steps may be excluded, i.e. the subject-matter consists in the inclusion of a stated member, integer or step or group of members, integers or steps. The terms “a” and “an” and “the” and similar reference used in the context of describing the invention (especially in the context of the claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. Recitation of ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”), provided herein is intended merely to better illustrate the invention and does not pose a limitation on the scope of the invention otherwise claimed. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the invention. 
     Several documents are cited throughout the text of this specification. Each of the documents cited herein (including all patents, patent applications, scientific publications, manufacturers specifications, instructions, etc.), whether supra or infra, are hereby incorporated by reference in their entirety. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention. 
     The term “human fibroblast growth factor 21 (FGF21)”, as used herein, refers to human wild-type FGF21 with the amino acid sequence of SEQ ID NO: 1 (also referred to as “full-length human wild-type FGF21”) or to naturally occurring variants thereof. The term “mature human wild-type FGF21” refers to human wild-type FGF21 lacking the natural signal sequence (also referred to as signal peptide), i.e., amino acids 1 to 28 (M1 to A28) of SEQ ID NO: 1, and is represented by SEQ ID NO: 2. Unless indicated otherwise, the numbering of particular amino acid residues in the FGF21 (poly-)peptides disclosed herein is in accordance with SEQ ID NO: 1. 
     The term “naturally occurring” when used in connection with biological materials, such as nucleic acid molecules, (poly-)peptides, host cells, and the like, refers to materials which are found in nature and are not manipulated by man. 
     In data bases open to the public like  The Universal Protein Resou rce (UniProt, online database available at uniprot.org) some naturally occurring variants of human FGF21 are described. Under the UniProtKB accession number Q9NSA1 two human FGF21 variants are listed: FGF21 G141→S (VAR_055375, SEQ ID NO: 172) and FGF21 L174→P (VAR_049064). The incidence for a proline at position 174 is more common than a leucine according to data bases like ExAC browser (P=73% versus L=27%; The Exome Aggregation Consortium; online available at exac.broadinstitute.org/variant/19-49261368-T-C) and AMP (Accelerating Medicines Partnership; online available at type2diabetesgenetics.org/variantInfo/variantInfo/rs739320). Thus, human FGF21 with P174 is referred to herein as human wild-type FGF21 (see SEQ ID NO: 1), whereas human FGF21 with L174 is included herein as a naturally occurring variant (see SEQ ID NO: 173). The mutation at position 174 has no impact on type 2 diabetes occurrence according to AMP. In addition, the mutation of L174 to P has no impact on the in vitro activity of FGF21 as assessed via In-Cell Western (described in Example 3). The EC50 of G-FGF21 with a proline in position 174 (SEQ ID NO: 3) is 0.24 nmol/L versus 0.21 nmol/L for G-FGF21 with a leucine in position 174 (SEQ ID NO: 174). The FGF21 variant G141␣S (VAR_055375, SEQ ID NO: 172) is very rare with an allele frequency below 0.1% (online available at exac.broadinstitute.org/variant/19-49261268-G-A) and was not further characterized in vitro. 
     A “variant of human fibroblast growth factor 21 (FGF21)” (also referred to as “FGF21 variant” or “FGF21 analogue”) in accordance with the present invention is a polypeptide which has a molecular structure which formally can be derived from the structure of human wild-type FGF21 or a naturally occurring variant thereof by deleting and/or substituting at least one amino acid residue in the naturally occurring human FGF21 (e.g., of SEQ ID NO: 1 or 2) and/or adding at least one amino acid residue. 
     In one embodiment, the variant of human FGF21 is a biologically active variant of human FGF21. The term “biologically active variant of human FGF21” refers to any variant that possesses an activity of the human wild-type FGF21 polypeptide, such as the ability to lower blood glucose, insulin, triglyceride, or cholesterol, reduce body weight, and/or to improve glucose tolerance, energy expenditure, or insulin sensitivity. Variants possessing a somewhat decreased level of FGF21 activity relative to the human wild-type FGF21 polypeptide can nonetheless be considered to be biologically active variants of human FGF21. 
     In some embodiments, a particular amino acid of the human wild-type FGF21 amino acid sequence (e.g., that of SEQ ID NO: 1 or 2) may be substituted by another amino acid. The term “another amino acid”, as used herein, typically relates to an amino acid which contributes to an increased stability, e.g., proteolytic and/or thermal stability, of the variant as compared to the human wild-type FGF21 (e.g., SEQ ID NO: 1 or 2). This may be achieved, for example, by the prevention of protease cleavage at or in proximity to the substituted amino acid or by formation of one or more additional disulfide bridges. 
     The term “amino acid” or “amino acid residue”, as used herein, refers to naturally occurring amino acids, unnatural amino acids, amino acid analogues and amino acid mimetics that function in a manner similar to the naturally occurring amino acids, all in their D and L stereoisomers if their structure allows such stereoisomeric forms. Amino acids are referred to herein by either their name, their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. 
     When used in connection with amino acids, the term “naturally occurring” refers to the 20 conventional amino acids (i.e., alanine (A), cysteine (C), aspartic acid (D), glutamic acid (E), phenylalanine (F), glycine (G), histidine (H), isoleucine (I), lysine (K), leucine (L), methionine (M), asparagine (N), proline (P), glutamine (Q), arginine (R), serine (S), threonine (T), valine (V), tryptophan (W), and tyrosine (Y)), as well as selenocysteine, pyrrolysine (PYL), and pyrroline-carboxylysine (PCL). 
     The term “unnatural amino acid”, as used herein, is meant to refer to amino acids that are not naturally encoded or found in the genetic code of any organism. They may, for example, be purely synthetic compounds. Examples of unnatural amino acids include, but are not limited to, hydroxyproline, gamma-carboxyglutamate, 0-phosphoserine, azetidinecarboxylic acid, 2-aminoadipic acid, 3-aminoadipic acid, beta-alanine, aminopropionic acid, 2-aminobutyric acid, 4-aminobutyric acid, 6-aminocaproic acid, 2-aminoheptanoic acid, 2-aminoisobutyric acid, 3-aminoisobutyric acid, 2-aminopimelic acid, tertiary-butylglycine, 2,4-diaminoisobutyric acid, desmosine, 2,2′-diaminopimelic acid, 2,3-diaminoproprionic acid, N-ethylglycine, N-methylglycine, N-ethylasparagine, homoproline, hydroxylysine, allo-hydroxylysine, 3-hydroxyproline, 4-hydroxyproline, isodesmosine, allo-isoleucine, N-methylalanine, N-methylglycine, N-methylisoleucine, N-methylpentylglycine, N-methylvaline, naphthalanine, norvaline, norleucine, ornithine, D-ornithine, D-arginine, p-aminophenylalanine, pentylglycine, pipecolic acid and thioproline. 
     The term “amino acid analogue”, as used herein, refers to compounds that have the same basic chemical structure as a naturally occurring amino acid. Amino acid analogues include the natural and unnatural amino acids which are chemically blocked, reversibly or irreversibly, or their C-terminal carboxy group, their N-terminal amino group and/or their side-chain functional groups are chemically modified. Such analogues include, but are not limited to, methionine sulfoxide, methionine sulfone, S-(carboxymethyl)-cysteine, S-(carboxymethyl)-cysteine sulfoxide, S-(carboxymethyl)-cysteine sulfone, aspartic acid-(betamethylester), N-ethylglycine, alanine carboxamide, homoserine, norleucine and methionine methyl sulfonium. 
     The term “amino acid mimetics”, as used herein, refers to chemical compounds that have a structure that is different from the general chemical structure of an amino acid, but functions in a manner similar to a naturally occurring amino acid. 
     The term “peptide”, as used herein, refers to a polymeric form of amino acids of any length, for example, comprising two or more, or 3 or more, or 4 or more, or 6 or more, or 8 or more, or 9 or more, or 10 or more, or 13 or more, or 16 or more, or 21 or more amino acids joined covalently by peptide bonds. The term “polypeptide” refers to large peptides, e.g., to peptides with more than 100 amino acid residues. The terms “polypeptide” and “protein” are used interchangeably herein. 
     In some embodiments, the variant comprises at least one additional amino acid at its N-terminus. In one embodiment, the at least one additional amino acid is selected from naturally occurring amino acids except proline, unnatural amino acids, amino acid analogues and amino acid mimetics. In one embodiment, the at least one additional amino acid is selected from the group consisting of G, A, N and C. In a particular embodiment, the at least one additional amino acid is G. 
     In some embodiments, the amino acid sequence of the variant according to the present invention has at least 90% or at least 91% or at least 92% or at least 93% or at least 94% or at least 95% or at least 96% or at least 97% or at least 98% sequence identity with one of SEQ ID NOs: 1 to 3, 172, 173 and 174, e.g., with mature human wild-type FGF21 (SEQ ID NO: 2). 
     “Sequence identity” between two amino acid sequences indicates the percentage of amino acids that are identical between the sequences. The optimal alignment of the sequences for comparison may be produced, besides manually, by means of the local homology algorithm of Smith and Waterman, 1981, Ads App. Math. 2, 482, by means of the local homology algorithm of Neddleman and Wunsch, 1970, J. Mol. Biol. 48, 443, by means of the similarity search method of Pearson and Lipman, 1988, Proc. Natl Acad. Sci. USA 85, 2444, or by means of computer programs which use these algorithms (GAP, BESTFIT, FASTA, BLAST P, BLAST N and TFASTA in Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Drive, Madison, Wis.). 
     The FGF21 variants featured in the invention typically have an increased proteolytic stability in human and/or murine blood plasma as compared to mature human wild-type FGF21 (SEQ ID NO: 2). 
     The term “proteolytic stability” refers to the ability of a (poly-)peptide to withstand proteolysis catalyzed by proteases. Examples of such proteases are serine proteases, threonine proteases, cysteine proteases, aspartate proteases and/or glutamic acid proteases. Examples of serine proteases are trypsin, chymotrypsin, plasmin, thrombin, granzyme and/or kallikreine. Examples of cysteine proteases are cathepsin K, caspase and/or calpain. According to the EC numbering such proteases are, e.g., classified according to the enzymatic activity as serine carboxypeptidases (EC 3.4.16), serine endopeptidases (EC 3.4.21), threonine endopeptidases (EC 3.4.25), cysteine carboxypeptidases (EC 3.4.18), cysteine endopeptidases (EC 3.4.22) or aspartate endopeptidases (EC 3.4.23). The term “proteolytic stability” may refer to in vitro or in vivo proteolytic stability. Means and methods to measure proteolytic stability of a given (poly-) peptide are known to a person skilled in the art. In one embodiment, proteolytic stability is measured with a sandwich immunoassay, e.g., essentially as described in Example 2. 
     The term “protease resistant peptide linker”, as used herein, refers to a peptide linker which has a low susceptibility to or withstands proteolysis catalyzed by proteases, e.g., proteases as described above. Such linker may have a length of 1 to 20 or 1 to 15 or 1 to 12 or 1 to 10 amino acids. In some embodiments, the linker may consist of as few as two amino acids (e.g., GA, GY, HH, GE or HE). In particular embodiments, the protease resistant peptide linker is of the peptide linker of SEQ ID NO:161, SEQ ID NO:162, SEQ ID NO:163, SEQ ID NO:164, SEQ ID NO:165, SEQ ID NO:166, SEQ ID NO:167, or SEQ ID NO:168. 
     The FGF21 variants featured in the invention will typically have an increased thermal stability as compared to mature human wild-type FGF21 (SEQ ID NO: 2). 
     The term “thermal stability” refers to the ability of a (poly-)peptide to resist irreversible change in its chemical or physical structure at higher temperatures, e.g., above 40° C. In particular embodiments, an increased thermal stability refers to an increased melting temperature (T M ) of the variant. Means and methods to measure thermal stability of a given (poly-)peptide are known to a person skilled in the art. In one embodiment, the thermal stability (e.g., Tn M ) is measured in a physiological buffer, such as PBS. In one embodiment, thermal stability (e.g., Tn M ) is measured with a thermal shift assay, e.g., essentially as described in Example 5. 
     In one embodiment, the melting temperature (Tn M ) of the variant is increased by at least 2° C., at least 4° C., at least 6° C., at least 8° C., at least 10° C., at least 12° C., at least 14° C., at least 16° C., at least 18° C. or at least 20° C. as compared to mature human wild-type FGF21 (SEQ ID NO: 2). In one embodiment, TM is increased by at least 18° C., e.g., between 18° C. and 30° C. or between 18° C. and 25° C., as compared to mature human wild-type FGF21 (SEQ ID NO: 2). 
     The biological function or activity of a FGF21 variant is usually measured and/or compared with human wild-type FGF21, e.g., mature human wild-type FGF21 (SEQ ID NO: 2), in a FGF21 activity assay generally known to a person skilled in the art. An FGF21 activity assay is, e.g., a “glucose uptake assay” as described in Kharitonenkov A. et al. (2005) J Clin Invest 115: 1627-1635. Alternatively, cellular assays, e.g., autophosporylation assays, such as an In-Cell Western (ICW) assay, can be used to measure the efficacy of the FGF21 variant to activate the FGF21 receptor of the FGF receptor:KLB complex or to stimulate the downstream intracellular signal transduction. 
     The term “In-Cell Western (ICW) assay”, as used herein, refers to an immunocytochemical assay, more particularly a quantitative immunofluorescence assay, usually performed in microplates (e.g., in a 96- or 384-well format). It combines the specificity of Western blotting with the reproducibility and throughput of ELISA (see, for example, Aguilar H. N. et al. (2010) PLoS ONE 5(4): e9965). Appropriate ICW assay systems are commercially available (e.g., from LI-COR Biosciences, USA). In one embodiment, the ICW assay is performed essentially as described in Example 3 or 4. 
     Typically, the FGF21 variants will induce phosphorylation of the mitogen-activated protein kinase (MAPK) ERK1/2. In one embodiment, the variants induce phosphorylation of the mitogen-activated protein kinase (MAPK) ERK1/2 with an EC50 of 100 nmol/L or lower, or 90 nmol/L or lower, or 80 nmol/L or lower, or 70 nmol/L or lower, or 60 nmol/L or lower, or 50 nmol/L or lower, or 40 nmol/L or lower, or 30 nmol/L or lower, or 20 nmol/L or lower, or 15 nmol/L or lower, or 12 nmol/L or lower, or 11 nmol/L or lower, or 10 nmol/L or lower, or 9 nmol/L or lower, or 8 nmol/L or lower, or 7 nmol/L or lower, or 6 nmol/L or lower, or 5 nmol/L or lower, or 4 nmol/L or lower, or 3 nmol/L or lower, or 2 nmol/L or lower, e.g., as determined in an In-Cell Western (ICW) assay. 
     A “label or tag allowing the detection and/or isolation of the variant” is meant to include any labels/tags known in the art for these purposes. Exemplary tags include affinity tags, such as chitin binding protein (CBP), maltose binding protein (MBP), glutathione-S-transferase (GST) and a His-tag, such as a histidine-rich sequence (e.g., SEQ ID NO: 5 or 6) or poly(His) (e.g., His 6 ); solubilization tags, such as thioredoxin (TRX) and poly(NANP); chromatography tags, such as a FLAG-tag; epitope tags, such as V5-tag, myc-tag and HA-tag; and fluorescent or luminescent labels or tags, such as fluorescent proteins (e.g., GFP, YFP, RFP etc.), fluorescent dyes and luciferase. 
     The amino acid sequence of a (poly)peptide label or tag may be introduced at any position within the amino acid sequence of the variant, and may, for example, take the shape of a loop within the encoded protein structure, or it may be N-terminally or C-terminally fused. In one embodiment, the label or tag is N-terminally fused. The label or tag may further contain a cleavage site (e.g., a TEV protease-cleavage site) that allows a removal of the label or tag from the variant. Similarly, non-peptidic labels or tags, e.g., fluorescent dyes, may be conjugated to the variant at any suitable site. 
     An FGF21 variant featured in the invention may also comprise an amino acid sequence for facilitating secretion of the molecule, such as an N-terminal secretion signal. In some embodiments, the secretion signal is a signal sequence that allows a sufficient passage through the secretory pathway and/or secretion into the extracellular environment. In some embodiments, the secretion signal sequence is cleavable and is removed from the variant. In one embodiment, the signal sequence is different from the natural signal sequence of human FGF21. The secretion signal sequence will be suitable for use in the cell or organism in which the variant is produced. In one embodiment, the secretion signal sequence comprises or consists of the amino acid sequence of SEQ ID NO: 4. 
     An FGF21 variant featured in the invention may further comprise a binding domain which serves, e.g., to enhance selectivity for a specific cell type or tissue. This can be achieved, e.g., by providing a binding domain that binds to a specific antigen expressed on the surface of said cell type or tissue. 
     An FGF21 variant may also be fused or conjugated to a half-life extension module. “Half-life” relates to the period of time which is needed to eliminate half of the activity, amount, or number of molecules, e.g., in vivo. Such modules are known to a person skilled in the art and include, for example, polymers (e.g., polyethylene glycol (PEG), hydroxyethyl starch (HES), hyaluronic acid, polysialic acid), unstructured (poly-)peptide chains, elastin-like polypeptides (ELPs), serum proteins (e.g., albumin, such as human serum albumin (HAS)), serum protein binding molecules (e.g., an albumin binding domain (ABD), an albumin binding fatty acid), antibodies, immunoglobulins, Fc regions/domains of immunoglobulins and immunoglobulin binding domains. 
     The term “unstructured (poly-)peptide chain”, as used herein, refers to a (poly-)peptide chain which lacks a fixed or ordered three-dimensional structure and is typically hydrophilic. Unstructured (poly-)peptide chains that extend the (in vivo) half-life of peptides and proteins they are fused to are known to a person skilled in the art and include, for example, XTEN (Schellenberger V. et al. (2009) Nat Biotechnol. 27(12): 1186-90) and PAS sequences 
     (Schlapschy M. et al. (2013) Protein Eng Des Sel. 26(8): 489-501). 
     The term “fused to”, as used herein, refers, in particular, to genetic fusion, e.g., by recombinant DNA technology. The amino acid sequence of a (poly)peptide half-life extension module may be introduced at any position within the amino acid sequence of the variant, and may, for example, take the shape of a loop within the encoded protein structure, or it may be N-terminally or C-terminally fused. 
     The term “conjugated to”, as used herein, refers, in particular, to chemical and/or enzymatic conjugation resulting in a stable covalent link between a (poly-)peptide and another molecule, e.g., the variant and the half-life extension module. Such conjugation may occur at the N- or C-terminus or at particular side chains of a (poly-)peptide, e.g., at lysine, cysteine, tyrosine or unnatural amino acid residues. 
     The term “fusion molecule” generally refers to molecules created by joining, in particular covalently linking, two or more distinct molecules (e.g., proteins and/or peptides) resulting in a single molecule with functional properties derived from each of the original molecules. In the case of proteins and/or peptides, the fusion molecule is referred to as “fusion protein”. Fusion molecules may be generated by genetic fusion (e.g., by recombinant DNA technology) or by chemical and/or enzymatic conjugation. The two or more distinct molecules may also be linked by suitable linker molecules, e.g., peptide linkers or non-peptidic polymers, such as polyethylene glycol (PEG). Peptide linkers may further comprise specific protease cleavage sites. 
     According to the present invention, the fusion molecule comprises at least one other active pharmaceutical ingredient in addition to the FGF21 variant. 
     The term “active pharmaceutical ingredient” (API), us used herein, includes any pharmaceutically active chemical or biological compound and any pharmaceutically acceptable salt thereof and any mixture thereof, that provides some pharmacologic effect and is used for treating or preventing a condition, e.g., a disease or disorder as defined herein. Exemplary pharmaceutically acceptable salts include hydrochloric, sulfuric, nitric, phosphoric, hydrobromic, maleric, malic, ascorbic, citric, tartaric, pamoic, lauric, stearic, palmitic, oleic, myristic, lauryl sulfuric, naphthalinesulfonic, linoleic, linolenic acid, and the like. As used herein, the terms “active pharmaceutical ingredient”, “active agent”, “active ingredient”, “active substance”, “therapeutically active compound” and “drug” are meant to be synonyms, i.e., have identical meaning. 
     In accordance with the present invention, an active pharmaceutical ingredient is optionally selected from:
         all drugs mentioned in the Rote Liste 2014, e.g. all antidiabetics mentioned in the Rote Liste 2014, chapter 12, all weight-reducing agents or appetite suppressants mentioned in the Rote Liste 2014, chapter 06, all lipid-lowering agents mentioned in the Rote Liste 2014, chapter 58, all antihypertensives mentioned in the Rote Liste 2014 chapter 17, all nephroprotectives mentioned in the Rote Liste, or all diuretics mentioned in the Rote Liste 2014, chapter 36;   insulin and insulin derivatives, for example: insulin glargine (e.g. Lantus®), higher than 100 U/mL concentrated insulin glargine, e.g. 270-330U/mL of insulin glargine or 300U/mL of insulin glargine (as disclosed in EP 2387989), insulin glulisine (e.g. Apidra®), insulin detemir (e.g. Levemir®), insulin lispro (e.g. Humalog®, Liprolog®), insulin degludec (e.g. DegludecPlus®, IdegLira (NN9068)), insulin aspart and aspart formulations (e.g. NovoLog®), basal insulin and analogues (e.g. LY2605541, LY2963016, NN1436), PEGylated insulin lispro (e.g. LY-275585), long-acting insulins (e.g. NN1436, Insumera (PE0139), AB-101, AB-102, Sensulin LLC), intermediate-acting insulins (e.g. Humulin®N, Novolin®N), fast-acting and short-acting insulins (e.g. Humulin®R, Novolin®R, Linjeta® (VlAject®), PH2O insulin, NN1218, HinsBet®), premixed insulins, SuliXen®, NN1045, insulin plus Symlin®, PE-0139, ACP-002 hydrogel insulin, and oral, inhalable, transdermal and buccal or sublingual insulins (e.g. Exubera®, Nasulin®, Afrezza®, insulin tregopil, TPM-02 insulin, Capsulin®, Oral-lyn®, Cobalamin® oral insulin, ORMD-0801, Oshadi oral insulin, NN1953, NN1954, NN1956, VlAtab®). also suitable are those insulin derivatives which are bonded to albumin or another protein by a bifunctional linker;   glucagon-like-peptide 1 (GLP-1), GLP-1 analogues, and GLP-1 receptor agonists, for example: GLP-1(7-37), GLP-1(7-36)amide, lixisenatide (e.g. Lyxumia®), exenatide (e.g. exendin-4, rExendin-4, Byetta®, Bydureon®, exenatide NexP), exenatide-LAR, liraglutide (e.g. Victoza®), semaglutide, taspoglutide, albiglutide, dulaglutide, albugon, oxyntomodulin, geniproside, ACP-003, CJC-1131, CJC-1134-PC, GSK-2374697, PB-1023, TTP-054, langlenatide (HM-11260C), CM-3, GLP-1 Eligen, AB-201, ORMD-0901, NN9924, NN9926, NN9927, Nodexen, Viador-GLP-1, CVX-096, ZYOG-1, ZYD-1, ZP-3022, CAM-2036, DA-3091, DA-15864, ARI-2651, ARI-2255, exenatide-XTEN (VRS-859), exenatide-XTEN+Glucagon-XTEN (VRS-859+AMX-808) and polymer-bound GLP-1 and GLP-1 analogues;   dual GLP-1/GIP agpmosts (e.g. RG-7697 (MAR-701), MAR-709, BHM081, BHM089, BHM098); dual GLP-1/glucagon receptor agonists (e.g. BHM-034, OAP-189 (PF-05212389, TKS-1225), TT-401/402, ZP2929, LAPS-HMOXM25, MOD-6030);   dual GLP-1/gastrin agonists (e.g. ZP-3022);   gastrointestinal peptides such as peptide YY 3-36 (PYY3-36) or analogues thereof and pancreatic polypeptide (PP) or analogues thereof;   glucagon receptor agonists or antagonists, glucose-dependent insulinotropic polypeptide (GIP) receptor agonists or antagonists, ghrelin antagonists or inverse agonists, xenin and analogues thereof;   dipeptidyl peptidase-IV (DPP-4) inhibitors, for example: alogliptin (e.g. Nesina®, Kazano®), linagliptin (e.g. Ondero®, Trajenta®, Tradjenta®, Trayenta®), saxagliptin (e.g. Onglyza® Komboglyze XR®), sitagliptin (e.g. Januvia®, Xelevia®, Tesavel®, Janumet®, Velmetia®, Juvisync®, Janumet XR®), anagliptin, teneligliptin (e.g. Tenelia®), trelagliptin, vildagliptin (e.g. Galvus®, Galvumet®), gemigliptin, omarigliptin, evogliptin, dutogliptin, DA-1229, MK-3102, KM-223, KRP-104, PBL-1427, Pinoxacin hydrochloride, and Ari-2243;   sodium-dependent glucose transporter 2 (SGLT-2) inhibitors, for example: canagliflozin, dapagliflozin, remogliflozin, remogliflozin etabonate, sergliflozin, empagliflozin, ipragliflozin, tofogliflozin, luseogliflozin, ertugliflozin, EGT-0001442, LIK-066, SBM-TFC-039, and KGA-3235 (DSP-3235);   dual inhibitors of SGLT-2 and SGLT-1 (e.g. LX-4211, LIK066).   SGLT-1 inhibitors (e.g. LX-2761, KGA-3235) or SGLT-1 inhibitors in combination with anti-obesity drugs such as ileal bile acid transfer (IBAT) inhibitors (e.g. GSK-1614235+GSK-2330672);   biguanides (e.g. metformin, buformin, phenformin);   thiazolidinediones (e.g. pioglitazone, rosiglitazone), glitazone analogues (e.g. lobeglitazone);   peroxisome proliferator-activated receptors (PPAR-)(alpha, gamma or alpha/gamma) agonists or modulators (e.g. saroglitazar (e.g. Lipaglyn®), GFT-505), or PPAR gamma partial agonists (e.g. Int-131);   sulfonylureas (e.g. tolbutamide, glibenclamide, glimepiride, Amaryl®, glipizide) and meglitinides (e.g. nateglinide, repaglinide, mitiglinide);   alpha-glucosidase inhibitors (e.g. acarbose, miglitol, voglibose);   amylin and amylin analogues (e.g. pramlintide, Symlin®);   G-protein coupled receptor 119 (GPR119) agonists (e.g. GSK-1292263, PSN-821, MBX-2982, APD-597, ARRY-981, ZYG-19, DS-8500, HM-47000, YH-Chem1);   GPR40 agonists (e.g. TUG-424, P-1736, P-11187, JTT-851, GW9508, CNX-011-67, AM-1638, AM-5262);   GPR120 agonists and GPR142 agonists;   systemic or low-absorbable TGR5 (GPBAR1=G-protein-coupled bile acid receptor 1) agonists (e.g. INT-777, XL-475, SB756050);   diabetes immunotherapeutics, for example: oral C-C chemokine receptor type 2 (CCR-2) antagonists (e.g. CCX-140, JNJ-41443532), interleukin 1 beta (IL-1β) antagonists (e.g. AC-201), or oral monoclonal antibodies (MoA) (e.g. methalozamide, VVP808, PAZ-320, P-1736, PF-05175157, PF-04937319);   anti-inflammatory agents for the treatment of the metabolic syndrome and diabetes, for example: nuclear factor kappa B inhibitors (e.g. Triolex®);   adenosine monophosphate-activated protein kinase (AMPK) stimulants, for example:       

     Imeglimin (PXL-008), Debio-0930 (MT-63-78), R-118;
         inhibitors of 11-beta-hydroxysteroid dehydrogenase 1 (11-beta-HSD-1) (e.g. LY2523199, BMS770767, RG-4929, BMS816336, AZD-8329, HSD-016, BI-135585);   activators of glucokinase (e.g. PF-04991532, TTP-399 (GK1-399), GKM-001 (ADV-1002401), ARRY-403 (AMC-151), TAK-329, TMG-123, ZYGK1);   inhibitors of diacylglycerol O-acyltransferase (DGAT) (e.g. pradigastat (LCQ-908)), inhibitors of protein tyrosine phosphatase 1 (e.g. trodusquemine), inhibitors of glucose-6-phosphatase, inhibitors of fructose-1,6-bisphosphatase, inhibitors of glycogen phosphorylase, inhibitors of phosphoenol pyruvate carboxykinase, inhibitors of glycogen synthase kinase, inhibitors of pyruvate dehydrogenase kinase;   modulators of glucose transporter-4, somatostatin receptor 3 agonists (e.g. MK-4256);   one or more lipid lowering agents are also suitable as combination partners, for example: 3-hydroxy-3-methylglutaryl-coenzym-A-reductase (HMG-CoA-reductase) inhibitors such as simvastatin (e.g. Zocor®, Inegy®, Simcor®), atorvastatin (e.g. Sortis®, Caduet®), rosuvastatin (e.g. Crestor®), pravastatin (e.g. Lipostat®, Selipran®), fluvastatin (e.g. Lescol®), pitavastatin (e.g. Livazo®, Livalo®), lovastatin (e.g. Mevacor®, Advicor®), mevastatin (e.g. Compactin®), rivastatin, cerivastatin (Lipobay®), fibrates such as bezafibrate (e.g. Cedur® retard), ciprofibrate (e.g. Hyperlipen®), fenofibrate (e.g. Antara®, Lipofen®, Lipanthyl®), gemfibrozil (e.g. Lopid®, Gevilon®), etofibrate, simfibrate, ronifibrate, clinofibrate, clofibride, nicotinic acid and derivatives thereof (e.g. niacin, including slow release formulations of niacin), nicotinic acid receptor 1 agonists (e.g.       

     GSK-256073), PPAR-delta agonists, acetyl-CoA-acetyltransferase (ACAT) inhibitors (e.g. avasimibe), cholesterol absorption inhibitors (e.g. ezetimibe, Ezetrol®, Zetia®, Liptruzet®, Vytorin®, S-556971), bile acid-binding substances (e.g. cholestyramine, colesevelam), ileal bile acid transport (IBAT) inhibitors (e.g. GSK-2330672, LUM-002), microsomal triglyceride transfer protein (MTP) inhibitors (e.g. lomitapide (AEGR-733), 
     SLx-4090, granotapide), modulators of proprotein convertase subtilisin/kexin type 9 (PCSK9) (e.g. alirocumab (REGN727/SAR236553), AMG-145, LGT-209, PF-04950615, MPSK3169A, LY3015014, ALD-306, ALN-PCS, BMS-962476, SPC5001, ISIS-394814, 1B20, LGT-210, 1D05, BMS-PCSK9Rx-2, SX-PCK9, RG7652), LDL receptor up-regulators, for example liver selective thyroid hormone receptor beta agonists (e.g. eprotirome (KB-2115), MB07811, sobetirome (QRX-431), VIA-3196, ZYT1), HDL-raising compounds such as: cholesteryl ester transfer protein (CETP) inhibitors (e.g. anacetrapib (MK0859), dalcetrapib, evacetrapib, JTT-302, DRL-17822, TA-8995, R-1658, LY-2484595, DS-1442), or dual CETP/PCSK9 inhibitors (e.g. K-312), ATP-binding cassette (ABC1) regulators, lipid metabolism modulators (e.g. BMS-823778, TAP-301, DRL-21994, DRL-21995), phospholipase A2 (PLA2) inhibitors (e.g. darapladib, Tyrisa®, varespladib, rilapladib), ApoA-I enhancers (e.g. RVX-208, CER-001, MDCO-216, CSL-112), cholesterol synthesis inhibitors (e.g. ETC-1002), lipid metabolism modulators (e.g. BMS-823778, TAP-301, DRL-21994, DRL-21995) and omega-3 fatty acids and derivatives thereof (e.g. icosapent ethyl (AMR101), Epanova®, AKR-063, NKPL-66, 
     PRC-4016, CAT-2003);
         bromocriptine (e.g. Cycloset®, Parlodel®), phentermine and phentermine formulations or combinations (e.g. Adipex-P, Ionamin, Qsymia®), benzphetamine (e.g. Didrex®), diethylpropion (e.g. Tenuate®), phendimetrazin (e.g. Adipost®, Bontril®), bupropion and combinations (e.g. Zyban®, Wellbutrin XL®, Contrave®, Empatic®), sibutramine (e.g. Reductil®, Meridia®), topiramat (e.g. Topamax®), zonisamid (e.g. Zonegran®), tesofensine, opioid antagonists such as naltrexone (e.g. Naltrexin®, naltrexone+bupropion), cannabinoid receptor 1 (CB1) antagonists (e.g. TM-38837), melanin-concentrating hormone (MCH-1) antagonists (e.g. BMS-830216, ALB-127158(a)), MC4 receptor agonists and partial agonists (e.g. AZD-2820, RM-493), neuropeptide Y5 (NPY5) or NPY2 antagonists (e.g. velneperit, S-234462), NPY4 agonists (e.g. PP-1420), beta-3-adrenergic receptor agonists, leptin or leptin mimetics, agonists of the 5-hydroxytryptamine 2c (5HT2c) receptor (e.g. lorcaserin, Belviq®), pramlintide/metreleptin, lipase inhibitors such as cetilistat (e.g. Cametor®), orlistat (e.g. Xenical®, Calobalin®), angiogenesis inhibitors (e.g. ALS-L1023), betahistidin and histamine H3 antagonists (e.g. HPP-404), AgRP (agouti related protein) inhibitors (e.g. TTP-435), serotonin re-uptake inhibitors such as fluoxetine (e.g. Fluctine®), duloxetine (e.g. Cymbalta®), dual or triple monoamine uptake inhibitors (dopamine, norepinephrine and serotonin re-uptake) such as sertraline (e.g. Zoloft®), tesofensine, methionine aminopeptidase 2 (MetAP2) inhibitors (e.g. beloranib), and antisense oligonucleotides against production of fibroblast growth factor receptor 4 (FGFR4) (e.g. ISIS-FGFR4Rx) or prohibitin targeting peptide-1 (e.g. Adipotide®);   nitric oxide donors, AT1 antagonists or angiotensin II (AT2) receptor antagonists such as telmisartan (e.g. Kinzal®, Micardis®), candesartan (e.g. Atacand®, Blopress®), valsartan (e.g. Diovan®, Co-Diovan®), losartan (e.g. Cosaar®), eprosartan (e.g. Teveten®), irbesartan (e.g. Aprovel®, CoAprovel®), olmesartan (e.g. Votum®, Olmetec®), tasosartan, azilsartan (e.g. Edarbi®), dual angiotensin receptor blockers (dual ARBs), angiotensin converting enzyme (ACE) inhibitors, ACE-2 activators, renin inhibitors, prorenin inhibitors, endothelin converting enzyme (ECE) inhibitors, endothelin receptor (ET1/ETA) blockers, endothelin antagonists, diuretics, aldosterone antagonists, aldosterone synthase inhibitors, alpha-blockers, antagonists of the alpha-2 adrenergic receptor, beta-blockers, mixed alpha-/beta-blockers, calcium antagonists, calcium channel blockers (CCBs), nasal formulations of the calcium channel blocker diltiazem (e.g. CP-404), dual mineralocorticoid/CCBs, centrally acting antihypertensives, inhibitors of neutral endopeptidase, aminopeptidase-A inhibitors, vasopeptide inhibitors, dual vasopeptide inhibitors such as neprilysin-ACE inhibitors or neprilysin-ECE inhibitors, dual-acting AT receptor-neprilysin inhibitors, dual AT1/ETA antagonists, advanced glycation end-product (AGE) breakers, recombinant renalase, blood pressure vaccines such as anti-RAAS (renin-angiotensin-aldosteron-system) vaccines, AT1- or AT2-vaccines, drugs based on hypertension pharmacogenomics such as modulators of genetic polymorphisms with antihypertensive response, thrombocyte aggregation inhibitors, and others or combinations thereof are suitable.       

     A “nucleic acid molecule” is deoxyribonucleic acid (DNA) or ribonucleic acid (RNA). A nucleic acid molecule may be in the form of a molecule which is single-stranded or double-stranded and linear or covalently closed to form a circle. 
     The term “DNA” relates to a molecule which comprises deoxyribonucleotide residues and s typically entirely or substantially composed of deoxyribonucleotide residues. “Deoxyribonucleotide” relates to a nucleotide which lacks a hydroxyl group at the 2′-position of a beta-D-ribofuranosyl group. The term “DNA” comprises isolated DNA such as partially or completely purified DNA, essentially pure DNA, synthetic DNA, and recombinantly generated 
     DNA and includes modified DNA which differs from naturally occurring DNA by addition, deletion, substitution and/or alteration of one or more nucleotides. Such alterations can include addition of non-nucleotide material, such as to the end(s) of a DNA or internally, for example at one or more nucleotides of the DNA. Nucleotides in DNA molecules can also comprise non-standard nucleotides, such as non-naturally occurring nucleotides or chemically synthesized nucleotides. These altered DNAs can be referred to as analogues or analogues of naturally-occurring DNA. When used in connection with nucleotides, the term “naturally occurring” refers to the bases adenine (A), cytosine (C), guanine (G), thymine (T), and uracil (U). 
     The term “RNA” relates to a molecule which comprises ribonucleotide residues and is typically entirely or substantially composed of ribonucleotide residues. “Ribonucleotide” relates to a nucleotide with a hydroxyl group at the 2′-position of a beta-D-ribofuranosyl group. The term “RNA” comprises isolated RNA such as partially or completely purified RNA, essentially pure RNA, synthetic RNA, and recombinantly generated RNA and includes modified RNA which differs from naturally occurring RNA by addition, deletion, substitution and/or alteration of one or more nucleotides. Such alterations can include addition of non-nucleotide material, such as to the end(s) of a RNA or internally, for example at one or more nucleotides of the RNA. Nucleotides in RNA molecules can also comprise non-standard nucleotides, such as non-naturally occurring nucleotides or chemically synthesized nucleotides or deoxynucleotides. These altered RNAs can be referred to as analogues or analogues of naturally-occurring RNA. 
     According to the invention, “RNA” refers to single-stranded RNA or double stranded RNA. In one embodiment, the RNA is mRNA, e.g., in vitro transcribed RNA (IVT RNA) or synthetic RNA. The RNA may also be modified, e.g., with one or more modifications increasing the stability (e.g., the half-life) of the RNA. Such modifications are known to a person skilled in the art and include, for example, 5′-caps or 5′cap analogues 
     The nucleic acid molecule according to the present invention may be contained/comprised in a vector. The term “vector”, as used herein, includes all vectors known to the skilled person, including plasmid vectors, cosmid vectors, phage vectors, such as lambda phage, viral vectors, such as adenoviral or baculoviral vectors, or artificial chromosome vectors such as bacterial artificial chromosomes (BAC), yeast artificial chromosomes (YAC), or P1 artificial chromosomes (PAC). Said vectors include expression as well as cloning vectors. Expression vectors comprise plasmids as well as viral vectors and generally contain a desired coding sequence and appropriate DNA sequences necessary for the expression of the operably linked coding sequence in a particular host organism (e.g., bacteria, yeast, plant, insect, or mammal) or in in vitro expression systems. Cloning vectors are generally used to engineer and amplify a certain desired DNA fragment and may lack functional sequences needed for expression of the desired DNA fragments. 
     Alternatively, the nucleic acid molecule according to the present invention may be integrated into a genome, e.g., the genome of a host cell. Means and methods to integrate a particular nucleic acid molecule into a genome are known to a person skilled in the art. 
     The term “cell” or “host cell” typically relates to an intact cell, i.e., a cell with an intact membrane that has not released its normal intracellular components such as enzymes, organelles, or genetic material. An intact cell is typically a viable cell, i.e. a living cell capable of carrying out its normal metabolic functions. The term typically relates to any cell which can be transfected or transformed with an exogenous nucleic acid. The cell when transfected or transformed with an exogenous nucleic acid and transferred to a recipient can typically express the nucleic acid in the recipient. The term “cell” includes prokaryotic cells, such as bacterial cells, and eukaryotic cells, such as yeast cells, fungal cells or mammalian cells. Suitable bacterial cells include cells from gram-negative bacterial strains, such as strains of  Escherichia coli, Proteus,  and  Pseudomonas,  and gram-positive bacterial strains, such as strains of  Bacillus, Streptomyces, Staphylococcus,  and  Lactococcus.  Suitable fungal cells include cells from the species of  Trichoderma, Neurospora,  and  Aspergillus.  Suitable yeast cells include cells from the species of  Saccharomyces  (for example,  Saccharomyces cerevisiae ),  Schizosaccharomyces  (for example,  Schizosaccharomyces pombe ), Pichia (for example,  Pichia pastoris  and  Pichia methanolica ), and  Hansenula.  Suitable mammalian cells include for example CHO cells, BHK cells, HeLa cells, COS cells, HEK293 and the like. In one embodiment, HEK293 cells are used. However, amphibian cells, insect cells, plant cells, and any other cells used in the art for the expression of heterologous proteins can be used as well. Mammalian cells, such as mammalian adipocyte cells, are particularly useful for adoptive transfer, including cells from humans, mice, hamsters, pigs, goats, and primates. The cells may be derived from a large number of tissue types and include primary cells and cell lines such as cells of the immune system, in particular antigen-presenting cells such as dendritic cells and T cells, stem cells such as hematopoietic stem cells and mesenchymal stem cells and other cell types. An antigen-presenting cell is a cell that displays antigen in the context of major histocompatibility complex on its surface. T cells may recognize this complex using their T cell receptor (TCR). The “cell” or “host cell” may be isolated or part of a tissue or organism, in particular a “non-human organism”. 
     The term “non-human organism”, as used herein, is meant to include non-human primates or other animals, in particular mammals, such as cows, horses, pigs, sheep, goats, dogs, cats, rabbits or rodents, such as mice, rats, guinea pigs and hamsters. 
     A pharmaceutical composition featured in the invention typically contains a therapeutically effective amount of a variant of human FGF21, optionally as part of a fusion molecule, the nucleic acid molecule or the host cell as described herein (also referred to herein as “agents”) to generate the desired reaction or the desired effect. 
     A pharmaceutical composition in accordance with the present invention may further comprise at least one other active pharmaceutical ingredient, e.g., at least one other active pharmaceutical ingredient as defined above. 
     A pharmaceutical composition in accordance with the present invention is typically a sterile composition. Pharmaceutical compositions are usually provided in a uniform dosage form and may be prepared in a manner known per se. A pharmaceutical composition may, e.g., be in the form of a solution or suspension. 
     A pharmaceutical composition may further comprise one or more carriers and/or excipients, all of which are pharmaceutically acceptable. The term “pharmaceutically acceptable”, as used herein, refers to the non-toxicity of a material which, does not interact with the action of the active agent of the pharmaceutical composition. 
     The term “carrier” refers to an organic or inorganic component, of a natural or synthetic nature, in which the active component is combined in order to facilitate, enhance or enable application. 
     According to the invention, the term “carrier” also includes one or more compatible solid or liquid fillers, diluents or encapsulating substances, which are suitable for administration to a subject. 
     Possible carrier substances for parenteral administration are, e.g., sterile water, Ringer&#39;s solution, Lactated Ringer&#39;s solution, physiological saline, bacteriostatic saline (e.g., saline containing 0.9% benzyl alcohol), phosphate-buffered saline (PBS), Hank&#39;s solution, polyalkylene glycols, hydrogenated naphthalenes and, in particular, biocompatible lactide polymers, lactide/glycolide copolymers or polyoxyethylene/polyoxy-propylene copolymers. 
     The term “excipient”, as used herein, is intended to include all substances which may be present in a pharmaceutical composition and which are not active ingredients, such as salts, binders (e.g., lactose, dextrose, sucrose, trehalose, sorbitol, mannitol), lubricants, thickeners, surface active agents, preservatives, emulsifiers, buffer substances, flavouring agents, or colorants. 
     Salts, which are not pharmaceutically acceptable, may be used for preparing pharmaceutically acceptable salts and are included in the invention. Pharmaceutically acceptable salts of this kind comprise in a non-limiting way those prepared from the following acids: hydrochloric, hydrobromic, sulfuric, nitric, phosphoric, maleic, acetic, salicylic, citric, formic, malonic, succinic acids, and the like. Pharmaceutically acceptable salts may also be prepared as alkali metal salts or alkaline earth metal salts, such as sodium salts, potassium salts or calcium salts. Salts may be added to adjust the ionic strength or tonicity. 
     Suitable preservatives for use in a pharmaceutical composition include antioxidants, citric acid, sodium citrate, benzalkonium chloride, chlorobutanol, cysteine, methionine, parabens and thimerosal. 
     Suitable buffer substances for use in a pharmaceutical composition include acetic acid in a salt, citric acid in a salt, boric acid in a salt and phosphoric acid in a salt. 
     The pharmaceutical composition may also be formulated as a stable lyophilized product that is reconstituted with an appropriate diluent, which, optionally, comprises one or more excipients as defined above. 
     As used herein, the term “kit of parts (in short: kit)” refers to an article of manufacture comprising one or more containers and, optionally, a data carrier. Said one or more containers may be filled with one or more of the above mentioned (re-)agents. Additional containers may be included in the kit that contain, e.g., diluents, buffers and further reagents. Said data carrier may be a non-electronical data carrier, e.g., a graphical data carrier such as an information leaflet, an information sheet, a bar code or an access code, or an electronical data carrier such as a compact disk (CD), a digital versatile disk (DVD), a microchip or another semiconductor-based electronical data carrier. The access code may allow the access to a database, e.g., an internet database, a centralized, or a decentralized database. Said data carrier may comprise instructions for the use of the FGF21 variants featured in the invention. 
     The agents and compositions described herein may be administered via any conventional route, e.g., orally, pulmonary, by inhalation or parenterally, including by injection or infusion. In one embodiment, parenteral administration is used, e.g., intravenously, intraarterially, subcutaneously, intradermally or intramuscularly. The agents and compositions described herein may also be administered through sustained release administration. 
     Pharmaceutical compositions suitable for parenteral administration usually comprise a sterile aqueous or non-aqueous preparation of the active compound, which is typically isotonic to the blood of the recipient. Examples of compatible carriers/solvents/diluents are sterile water, Ringer&#39;s solution, Lactated Ringer&#39;s solution, physiological saline, bacteriostatic saline (e.g., saline containing 0.9% benzyl alcohol), phosphate-buffered saline (PBS) and Hank&#39;s solution. In addition, usually sterile, fixed oils may be used as solution or suspension medium. 
     The agents and compositions described herein are usually administered in therapeutically effective amounts. A “therapeutically effective amount” refers to the amount, which achieves a desired therapeutic reaction or a desired therapeutic effect alone or together with further doses, typically without causing unacceptable side-effects. In the case of treatment of a particular disease or of a particular condition, the desired reaction typically relates to inhibition of the course of the disease. This comprises slowing down the progress of the disease and, in particular, interrupting or reversing the progress of the disease. The desired reaction in a treatment of a disease or of a condition may also be delay of the onset or a prevention of the onset of said disease or said condition. An effective amount of an agent or composition described herein will depend on the condition to be treated, the severeness of the disease, the individual parameters of the subject, including age, physiological condition, size and weight, the duration of treatment, the type of an accompanying therapy (if present), the specific route of administration and similar factors. Accordingly, the doses administered of the agents described herein may depend on various of such parameters. In the case that a reaction in a subject is insufficient with an initial dose, higher doses (or effectively higher doses achieved by a different, more localized route of administration) may be used. 
     The present invention also provides a combination of a variant of human FGF21 of the present invention with at least one other active pharmaceutical ingredient. 
     In one embodiment, the combination of the variant of human FGF21 of the present invention with at least one other active pharmaceutical ingredient can be applied either by separate administration of the active pharmaceutical ingredient to the patient or in the form of combination products in which a plurality of active pharmaceutical ingredients are present in one pharmaceutical composition. When administered separately, administration may occur simultaneously or sequentially, in any order. The amount of the FGF21 variant and the other active pharmaceutical ingredient(s) and the relative timings of administration will be selected in order to achieve the desired combined therapeutic effect. The administration of the combination may be concomitantly in: (1) a unitary pharmaceutical composition including all active pharmaceutical ingredient; or (2) separate pharmaceutical compositions each including at least one of the active pharmaceutical ingredient. Alternatively, the combination may be administered separately in a sequential manner wherein one treatment agent is administered first and the other second or vice versa. Such sequential administration may be close in time or remote in time. 
     According to the invention, the term “disease or disorder” refers to any pathological or unhealthy state, in particular obesity, overweight, metabolic syndrome, diabetes mellitus, hyperglycemia, dyslipidemia, non-alcoholic steatohepatitis (NASH) and/or atherosclerosis. 
     The term “obesity” refers to a medical condition in which excess body fat has accumulated to the extent that it may have a negative effect on health. In terms of a human (adult) subject, obesity can be defined as a body mass index (BMI) greater than or equal to 30 kg/m 2  (BMI≥30 kg/m 2 ). 
     The term “overweight” refers to a medical condition in which the amount of body fat is higher than is optimally healthy. In terms of a human (adult) subject, obesity can be defined as a body mass index (BMI) greater than or equal to 25 kg/m 2  (e.g., 25 kg/m 2 ≤BMI&lt;30 kg/m 2 ). 
     The BMI is a simple index of weight-for-height that is commonly used to classify overweight and obesity in adults. It is defined as a person&#39;s weight in kilograms divided by the square of his/her height in meters (kg/m 2 ). 
     “Metabolic syndrome” can be defined as a clustering of at least three of the following medical conditions: abdominal (central) obesity (e.g., defined as waist circumference≥94 cm for Europid men and ≥80 cm for Europid women, with ethnicity specific values for other groups), elevated blood pressure (e.g., 130/85 mmHg or higher), elevated fasting plasma glucose (e.g., at least 100 mg/dL), high serum triglycerides (e.g., at least 150 mg/dL), and low high-density lipoprotein (HDL) levels (e.g., less than 40 mg/dL for males and less than 50 mg/dL for females). 
     “Diabetes mellitus” (also simply referred to as “diabetes”) refers to a group of metabolic diseases characterized by high levels of blood glucose resulting from defects in insulin production, insulin action, or both. In one embodiment, diabetes mellitus is selected from the group consisting of type 1 diabetes mellitus, type 2 diabetes mellitus, gestational diabetes mellitus, late onset autoimmune diabetes in the adult (LADA), maturity onset diabetes of the young (MODY) and other types of diabetes resulting from specific genetic conditions, drugs, malnutrition, infections and other illnesses. 
     The current WHO diagnostic criteria for diabetes mellitus are as follows: fasting plasma glucose ÷7.0 mmol/l (126 mg/dL) or 2-h plasma glucose ≥11.1 mmol/l (200 mg/dL). 
     “Type 1 diabetes mellitus” (also known as “insulin-dependent diabetes (IDDM)” or “juvenile diabetes”) is a condition characterized by high blood glucose levels caused by total lack of insulin. This occurs when the body&#39;s immune system attacks the insulin producing beta cells in the pancreas and destroys them. The pancreas then produces little or no insulin. Pancreatic removal or disease may also lead to loss of insulin-producing beta cells. Type 1 diabetes mellitus accounts for between 5% and 10% of cases of diabetes. 
     “Type 2 diabetes mellitus” (also known as “non-insulin-dependent diabetes (NIDDM)” or “adult-onset diabetes”) is a condition characterized by excess glucose production in spite of the availability of insulin, and circulating glucose levels remain excessively high as a result of inadequate glucose clearance (insulin action). Type 2 diabetes mellitus may account for about 90% to 95% of all diagnosed cases of diabetes. 
     “Gestational diabetes” is a condition in which women without previously diagnosed diabetes exhibit high blood glucose levels during pregnancy (especially during the third trimester). Gestational diabetes affects 3-10% of pregnancies, depending on the population studied. 
     “Late onset autoimmune diabetes in the adult (LADA)” (also referred to as “slow onset type 1 diabetes”) is a form of type 1 diabetes mellitus that occurs in adults, often with a slower course of onset. 
     “Maturity onset diabetes of the young (MODY)” refers to a hereditary form of diabetes caused by mutations in an autosomal dominant gene disrupting insulin production. 
     The term “hyperglycemia” refers to an excess of sugar (glucose) in the blood. 
     The term “dyslipidemia” refers to a disorder of lipoprotein metabolism, including lipoprotein overproduction (“hyperlipidemia”) or deficiency (“hypolipidemia”). Dyslipidemias may be manifested by elevation of the total cholesterol, low-density lipoprotein (LDL) cholesterol and/or triglyceride concentrations, and/or a decrease in high-density lipoprotein (HDL) cholesterol concentration in the blood. 
     Non-alcoholic steatohepatitis (NASH) is a liver disease characterized by an accumulation of fat (lipid droplets), along with inflammation and degeneration of hepatocytes. Once installed, the disease is accompanied with a high risk of cirrhosis, a state where the liver functions are altered and can progress to liver insufficiency. Thereafter, NASH often progresses to liver cancer. 
     “Atherosclerosis” is a vascular disease characterized by irregularly distributed lipid deposits called plaque in the intima of large and medium-sized arteries that may cause narrowing of arterial lumens and proceed to fibrosis and calcification. Lesions are usually focal and progress slowly and intermittently. Occasionally plaque rupture occurs leading to obstruction of blood flow resulting in tissue death distal to the obstruction. Limitation of blood flow accounts for most clinical manifestations, which vary with the distribution and severity of the obstruction. 
     The term “medicament”, as used herein, refers to a substance/composition used in therapy, i.e., in the treatment of a disease or disorder. 
     By “treat” is meant to administer a compound or composition or a combination of compounds or compositions to a subject in order to prevent or eliminate a disease or disorder; arrest or slow a disease or disorder in a subject; inhibit or slow the development of a new disease or disorder in a subject; decrease the frequency or severity of symptoms and/or recurrences in a subject who currently has or who previously has had a disease or disorder; and/or prolong, i.e., increase, the lifespan of the subject. 
     In particular, the term “treating/treatment of a disease or disorder” includes curing, shortening the duration, ameliorating, preventing, slowing down or inhibiting progression or worsening, or preventing or delaying the onset of a disease or disorder or the symptoms thereof. 
     The term “subject” means according to the invention a subject for treatment, in particular a diseased subject (also referred to as “patient”), including human beings, non-human primates or other animals, in particular mammals, such as cows, horses, pigs, sheep, goats, dogs, cats, rabbits or rodents, such as mice, rats, guinea pigs and hamsters. In one embodiment, the subject/patient is a human being. 
     FURTHER REFERENCES 
     Tomlinson E. et al. (2002) Endocrinology 143: 1741-1747; 
     Holt J. A. et al. (2003) Genes Dev 17: 1581-1591; 
     Shimada T. et al. (2004) J Bone Miner Res 19: 429-435; 
     Inagaki T. et al. (2005) Cell Metab 5: 415-425; 
     Lundasen T. et al. (2006) J Intern Med 260: 530-536; 
     Nishimura T. et al. (2000) Biochim Biophys Acta 1492: 203-206; 
     Kharitonenkov A. et al. (2005) J Clin Invest 115: 1627-1635; 
     Kharitonenkov A. et al. (2007) Endocrinology 148: 774-781; 
     Huang X. et al. (2006) Mol Carcinog 45: 934-942; 
     Wente W. et al. (2006) Diabetes 55: 2470-2478; 
     Ornitz D. M. et al. (2001) Genome Biol 2: REVIEWS3005; 
     Nicholes K. et al. (2002) Am J Pathol 160: 2295-2307; 
     Eswarakumar V. P. et al. (2005) Cytokine Growth Factor Rev 16: 139-149; 
     Dostalova I. et al. (2009) Physiol Res 58: 1-7; 
     WO 2003/011213 A2 and US 2004/0259780 A1; 
     WO 2005/061712 A1 and US 2007/0142278 A1; 
     WO 2006/028595 A2 and US 2008/0103096 A1; 
     WO 2006/028714 Al and US 2007/0299007 A1; 
     WO 2006/065582 A2 and US 2007/0293430 A1; 
     WO 2008/121563 A2 and US 2008/0255045 A1. 
     EXAMPLES 
     Example 1: Expression of FGF21 Variants in HEK293 Cells or in  Escherichia coli  ( E. coli ) 
     The FGF21 variants were produced by transient transfection of HEK293 cells or in  E. coli.    
     (a) Expression of FGF21 variants in HEK293 cells 
     The DNA sequence of the FGF21 variants was N-terminally fused to an IL2 signal sequence (SEQ ID NO: 4) followed by a Histidine-rich sequence (His-tag) and a TEV protease-cleavage site (SEQ ID NO: 5 or 6). The signal sequence was required for secretion of the desired proteins into the culture medium. The desired proteins were purified from the culture supernatant using immobilized metal-ion affinity chromatography (IMAC) (cOmplete His-Tag Purification Column, Roche). After elution from the IMAC-column, the N-terminal His-tag was optionally cleaved by addition of TEV protease. After His-tag cleavage, the cleavage reaction solution was passed a second time over an Immobilized-Metal Affinity Column (IMAC-column) (cOmplete His-Tag Purification Column, Roche), collecting the (His-tag-free) flow-through fraction. The protein was further purified using a gelfiltration column with phosphate buffered saline (PBS, Gibco) as running buffer. Fractions containing the desired proteins were collected, pooled, concentrated and stored at −80° C. until further usage. 
     (b) Expression of FGF21 Variants in  E. coli    
     The FGF21 proteins of SEQ ID NO: 2 (mature human wild-type FGF21, i.e. without signal sequence; also referred to as FGF21 H29-5209) and SEQ ID NO: 3 (mature human wild-type FGF21 with an additional N-terminal G; also referred to as G+FGF21 H29-5209 or G-FGF21 H29-5209 or simply G-FGF21) as well as the proteins of SEQ ID NOs: 124 through SEQ ID NO:157 and SEQ ID NO: 174 were expressed in E. coll. The DNA sequences of the FGF21 proteins were N-terminally fused to a Histidine-rich sequence (His-tag) and a TEV or SUMO protease-cleavage site (SEQ ID NO: 6 or 7). The desired proteins were purified using immobilized metal-ion affinity chromatography (IMAC) (His Trap HP, GE Healthcare) followed by cleavage of the N-terminal His-tag by addition of TEV or SUMO protease. After His-tag cleavage, the cleavage reaction solution was purified using an ion exchange column, followed by a gelfiltration step using phosphate buffered saline (PBS, Gibco) as running buffer. Fractions containing the desired proteins were collected, pooled, concentrated and stored at −80° C. until further usage. 
     Example 2: Stability Measurement of FGF21 Variants in Human and Murine Plasma 
     In vivo pharmacokinetic (PK) analysis of mature human wild-type FGF21 in mice and rat revealed a half-life of less than 1 hour due to a rapid clearance via the kidney (Kharitonenkov A. et al. (2007) Endocrinology 148(2): 774-781; Stein S. et al. (2009) Diabetes Care 32(1): 126-8; Lin Z. et al. (2011) PLoS ONE 6(4): e18398). In addition, FGF21 was also found to be susceptible to in vivo proteolytic degradation. This degradation negatively affects the therapeutic potential of FGF21 significantly (Table 1) (Hecht R. et al. (2012) PLoS ONE 7(11): e49345; Micanovic R. et al. (2009) J Cell Physiol. 219(2): 227-34; Yie J. et al. (2012) Chem Biol Drug Des. 79(4): 398-410). 
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 Loss of C-terminal amino acids of FGF21 decreases its in vitro activity. 
               
            
           
           
               
               
               
            
               
                   
                   
                 pFGFR ICW 
               
            
           
           
               
               
               
               
            
               
                   
                 Mw 
                 EC50  
                 Emax  
               
               
                 Protein 
                 (kDa) 
                 (nmol/L) 
                 (%) 
               
               
                   
               
               
                 FGF21, mature human wild-type  
                 19.393 
                 4.49 
                 146 
               
               
                 (H29-S209) 
                   
                   
                   
               
               
                 FGF21 truncated C-terminus 
                 17.973 
                 inactive 
                 inactive 
               
               
                 delta15 amino acids (H29-L194) 
                   
                   
                   
               
               
                 FGF21 truncated C-terminus  
                 16.234 
                 inactive 
                 inactive 
               
               
                 delta33 amino acids (H29-E176) 
               
               
                   
               
            
           
         
       
     
     To learn more about the metabolic stability of human FGF21 and to identify metabolites formed by proteolytic degradation in blood the protein was analysed as described subsequently. In detail, 15 μmol/L of human wild-type FGF21 (SEQ ID NO: 3) was incubated in vitro in human, mouse, and rat lithium heparin plasma at 37° C. for 60 minutes. At the end of incubation, samples were worked up with Norgen Serum Depletion Kit (Norgen Biotek, Thorold, ON, Canada, #17300) according to the procedure described by the manufacturer. Major serum proteins including albumin, alpha-antitrypsin, transferrin and haptoglobin were depleted from plasma samples. At 4° C. 500 pL plasma samples were applied to the activated spin column and centrifuged in a benchtop microcentrifuge at 6,700 x g for one minute, wherein the flow-through was discarded. The column was washed two times by addition of 500 μL Wash Buffer and centrifugation for one minute. Two times 100 μL Elution Buffer were added to the column and centrifuged for one minute to elute bound proteins. 
     Metabolites were identified by liquid chromatography-mass spectrometry (LC-MS) experiments on an ion trap mass spectrometer within the mass range from m/z 650 to m/z 1700 in the positive ionization mode with accurate mass measurement (Thermo-Fisher LTQ-Orbitrap, NanoLC 1D Ultra, AS2 Autosampler). The HPLC conditions used for metabolite identification were the following: column Vydac MS C4 300 A, 5 pm, 150 mm x 75 pm (Grace, Lokeren, Belgium); mobile phase: eluent A 0.1% Formic acid solution with 2% (v/v) of acetonitrile and eluent B Acetonitrile+0.1% Formic acid; run time 75 min; flow rate: 280 nL/min. Multi charged masses were deconvoluted by Magtran software (Zhongqi Zhang), and the protein sequence of the formed metabolite was elucidated by GPMAW program (Lighthouse Data, Odense, Denmark). 
     Incubation of FGF21 of SEQ ID NO: 3 for 1 hour as described above in blood plasma resulted in a rapid cleavage, and several metabolites could be detected which are listed in Table 2. A cleavage product corresponding to the amino acid sequence from position 29 to position 199 of human wild-type FGF21 (SEQ ID NO: 1) was formed in the plasma of all species investigated. This metabolite can be described as human-FGF21-des-decapeptide, having a molecular weight of 18428.56 Da. Mass spectra and deconvolution of human FGF21 of SEQ ID NO: 3 and its cleavage products are presented in  FIGS. 1A-1F . 
     
       
         
           
               
             
               
                 TABLE 2 
               
             
            
               
                   
               
               
                 Metabolites of human FGF21 identified in plasma of human, rat, 
               
               
                 and mice after 1 hour incubation (numbering of the amino 
               
               
                 acids refers to human wild-type FGF21 of SEQ ID NO: 1).  
               
            
           
           
               
               
               
               
               
            
               
                 Input 
                 Found 
                 Dev. 
                 from 
                 to 
               
               
                   
               
            
           
           
               
            
               
                 Human 
               
            
           
           
               
               
               
               
               
            
               
                 18429 
                 18428.56 
                 24 
                 H29  
                 P199 
               
               
                 19392 
                 19392.55 
                 28 
                 H29  
                 S209 
               
               
                 5113.7 
                 5113.76 
                 12 
                 R159 
                 S209 
               
               
                 5379 
                 5379.07 
                 13 
                 P156 
                 S209 
               
               
                 5379 
                 5379.08 
                 14 
                 P152 
                 S204 
               
               
                 5379 
                 5379.08 
                 14 
                 H153 
                 P205 
               
               
                 5379 
                 5379.08 
                 14 
                 S151 
                 R203 
               
               
                 4732.4 
                 4732.33 
                 15 
                 R163 
                 S209 
               
            
           
           
               
            
               
                 Rat 
               
            
           
           
               
               
               
               
               
            
               
                 18428 
                 18428.56 
                 30 
                 H29  
                 P199 
               
               
                 5113.7 
                 5113.76 
                 12 
                 R159 
                 S209 
               
            
           
           
               
            
               
                 Mouse 
               
            
           
           
               
               
               
               
               
            
               
                 18427 
                 18428.56 
                 85 
                 H29  
                 P199 
               
               
                 8564.67 
                 8564.73 
                 7 
                 L80  
                 P156 
               
               
                 8564.67 
                 8564.46 
                 25 
                 Y48  
                 E125 
               
               
                 8564.67 
                 8565.54 
                 102 
                 Y111 
                 S190 
               
               
                 8564.67 
                 8565.64 
                 113 
                 V90  
                 P166 
               
               
                   
               
            
           
         
       
     
     The in vitro stability of mature human wild-type FGF21 (SEQ ID NO: 2) and various FGF21 variants was analyzed by incubating the proteins for up to 24 hours at 37° C. in human or mouse blood plasma, and analysis of the remaining intact, full length FGF21 protein using a sandwich immunoassay. In detail, mature human FGF21 (SEQ ID NO: 2) or variants were added to blood samples (K2EDTA plasma, mixture of several healthy human donors) at a concentration of 100 ng/mL and incubated for up to 24 hours at 2×concentrated protease inhibitor (Protease Inhibitor Cocktail Tablets, Roche). The samples were then transferred to −80° C. until further processing. Subsequently, the amount of remaining intact, full length FGF21 protein was assessed using Human Intact FGF21 ELISA (F2131-K01, Eagle Biosciences, Nashua, NH, USA). The assay utilizes the sandwich technique with the two selected antibodies specifically binding to the N-terminus of mature human FGF21 (aa1-7) and the C-terminus (aa175-181) as described by the vendor which corresponds to amino aa29-35 and aa203-209 of full-length human wild-type FGF21 (SEQ ID NO: 1). The concentration of each individual sample at start of the incubation was used as internal reference and correlated to concentrations measured at later time points. 
     The results of the tested recombinant FGF21 variants for stability in human and murine plasma are summarized in Table 3, and representative results are shown in  FIG. 2 . 
     
       
         
           
               
             
               
                 TABLE 3  
               
             
            
               
                   
               
               
                 Influence of FGF21 mutations on its stability 
               
               
                 against proteolysis (numbering of the namino acids 
               
               
                 refers to human wild-type FGF21 of SEQ ID NO: 1). 
               
            
           
           
               
               
            
               
                   
                 Stability  
               
               
                   
                 against 
               
               
                 Protein/Mutation 
                 Proteolysis 
               
               
                   
               
               
                 FGF21, human (SEQ ID NO: 2) 
                 Reference 
               
               
                   
               
               
                 G-FGF21 (SEQ ID NO: 3) 
                 Reference 
               
               
                   
               
               
                 FGF21 V197D, G198V, P199D, S200H 
                 increased 
               
               
                 (SEQ ID NO: 26) 
                   
               
               
                   
               
               
                 FGF21 V197-R203 deleted and replaced 
                 not increased 
               
               
                 by GHRSHLQTVF (SEQ ID NO: 27) 
                   
               
               
                   
               
               
                 FGF21 P199S, S200P (SEQ ID NO: 9) 
                 increased 
               
               
                   
               
               
                 FGF21 P199Q, Q201P (SEQ ID NO: 10) 
                 increased 
               
               
                   
               
               
                 FGF21 P199 deleted (SEQ ID NO: 11) 
                 increased 
               
               
                   
               
               
                 FGF21 M196P, V197G, G198V, P199M 
                 increased 
               
               
                 (SEQ ID NO: 12) 
                   
               
               
                   
               
               
                 FGF21 M196V, V197M (SEQ ID NO: 13) 
                 not increased 
               
               
                   
               
               
                 FGF21 V197E (SEQ ID NO: 14) 
                 not increased 
               
               
                   
               
               
                 FGF21 G198E (SEQ ID NO: 16) 
                 increased 
               
               
                   
               
               
                 FGF21 G198D (SEQ ID NO: 17) 
                 increased 
               
               
                   
               
               
                 FGF21 G198R (SEQ ID NO: 18) 
                 increased 
               
               
                   
               
               
                 FGF21 G198K (SEQ ID NO: 19) 
                 increased 
               
               
                   
               
               
                 FGF21 G198Y (SEQ ID NO: 20) 
                 increased 
               
               
                   
               
               
                 FGF21 P199G (SEQ ID NO: 21) 
                 increased 
               
               
                   
               
               
                 FGF21 P199T (SEQ ID NO: 23) 
                 increased 
               
               
                   
               
               
                 FGF21 G198P, P199G (SEQ ID NO: 24) 
                 increased 
               
               
                   
               
               
                 FGF21 S200Q, Q2015 (SEQ ID NO: 25) 
                 not increased 
               
               
                   
               
               
                 FGF21 S195C, S200C (SEQ ID NO: 8) 
                 not increased 
               
               
                   
               
               
                 FGF21 GGGGS inserted between G198 
                 not increased 
               
               
                 and P199 (SEQ ID NO: 15) 
                   
               
               
                   
               
               
                 FGF21 P199R (SEQ ID NO: 22) 
                 increased 
               
               
                   
               
            
           
         
       
     
     Example 3: In Vitro Cellular Assay for Human FGF21 Receptor Rfficacy in CHO Cells (In-Cell Western) 
     The cellular in vitro efficacy of mature human FGF21 (SEQ ID NO: 2) or FGF21 variants was measured using a specific and highly sensitive In-Cell Western (ICW) assay. The ICW assay is an immunocytochemical assay usually performed in microplate format. CHO Flp-In cells (Invitrogen, Darmstadt, Germany) stably expressing the human FGFR1c together with human beta-Klotho (KLB) were used for a FGF21 receptor autophosphorylation assay using In-Cell Western (Aguilar H. N. et al. (2010) PLoS ONE 5(4): e9965). In order to determine the receptor autophosphorylation level or downstream activation of the MAP kinase ERK1/2, 2×10 4  cells/well were seeded into 96-well plates and grown for 48 h. Cells were serum starved with serum-free medium Ham&#39;s F-12 Nutrient Mix with GlutaMAX (Gibco, Darmstadt, Germany) for 3-4 h. The cells were subsequently treated with increasing concentrations of either mature human FGF21 (SEQ ID NO: 2), the indicated FGF21 variant, or other peptides for 5 min at 37° C. After incubation, the medium was discarded, and the cells were fixed in 3.7% freshly prepared para-formaldehyde for 20 min. Cells were permeabilized with 0.1% Triton-X-100 in PBS for 20 min. 
     Blocking was performed with Odyssey blocking buffer (LICOR, Bad Homburg, Germany) for 2 h at room temperature. As primary antibody, anti-pFGFR Tyr653/654 (New England Biolabs, Frankfurt, Germany) or anti-pERK Phospho-p44/42 MAP Kinase Thr202/Tyr204 (Cell Signaling) was added and incubated overnight at 4° C. After incubation of the primary antibody, cells were washed with PBS plus 0.1% Tween20. The secondary anti-Mouse 800CW antibody (LICOR, Bad Homburg, Germany) was incubated for 1 h at room temperature. Subsequently, cells were washed again with PBS plus 0.1% Tween20, and infrared dye signals were quantified with an Odyssey imager (LICOR, Bad Homburg, Germany). Results were normalized by quantification of DNA with TO-PRO3 dye (Invitrogen, Karlsruhe, Germany). Data were obtained as arbitrary units (AU), and EC50 values were obtained from dose-response curves and are summarized in 
     Table 4.  FIGS. 3A -3W show the results from an ICW with CHO cells overexpressing human FGFR1c plus KLB. 
     
       
         
           
               
             
               
                 TABLE 4  
               
             
            
               
                   
               
               
                 EC50-values of mature human FGF21 (SEQ ID NO: 2) 
               
               
                 and FGF21 variants measured via ICW pERK in CHO cells. 
               
               
                 Unless otherwise specified, tested FGF21 proteins were 
               
               
                 purified by using an N-terminal tag (SEQ ID NO: 5 or 6), 
               
               
                 i.e. unless otherwise specified the FGF21 proteins carried  
               
               
                 an N-terminal tag when determining the EC50-values. 
               
               
                 The numbering of the amino acids refers 
               
               
                 to human wild-type FGF21 of SEQ ID NO: 1. 
               
            
           
           
               
               
            
               
                   
                 pERK ICW 
               
               
                 Protein/Mutation 
                 EC50 (nmol/L) 
               
               
                   
               
            
           
           
               
               
            
               
                 FGF21, human (SEQ ID NO: 2), without N-terminal 
                 0.17 
               
               
                 tag 
                   
               
               
                   
               
               
                 G-FGF21 (SEQ ID NO: 3), without N-terminal tag 
                 0.24 
               
               
                   
               
               
                 FGF21, human (SEQ ID NO: 2) 
                 1.03 
               
               
                   
               
               
                 FGF21 V197D, G198V, P199D, S200H (SEQ ID NO: 
                 85.84 
               
               
                 26) 
                   
               
               
                   
               
               
                 FGF21 V197-R203 deleted and replaced by 
                 6.08 
               
               
                 GHRSHLQTVF (SEQ ID NO: 27) 
                   
               
               
                   
               
               
                 FGF21 V197-R203 deleted and replaced by 
                 78.10 
               
               
                 GLNSMV (SEQ ID NO: 28) 
                   
               
               
                   
               
               
                 FGF21 P199S, S200P (SEQ ID NO: 9) 
                 1.72 
               
               
                   
               
               
                 FGF21 P199Q, Q201P (SEQ ID NO: 10) 
                 1.76 
               
               
                   
               
               
                 FGF21 P199 deleted (SEQ ID NO: 11) 
                 2.31 
               
               
                   
               
               
                 FGF21 M196P, V197G, G198V, P199M (SEQ ID 
                 inactive 
               
               
                 NO: 12) 
                   
               
               
                   
               
               
                 FGF21 M196V, V197M (SEQ ID NO: 13) 
                 26.73 
               
               
                   
               
               
                 FGF21 V197E (SEQ ID NO: 14) 
                 31.45 
               
               
                   
               
               
                 FGF21 G198E (SEQ ID NO: 16) 
                 4.53 
               
               
                   
               
               
                 FGF21 G198D (SEQ ID NO: 17) 
                 4.94 
               
               
                   
               
               
                 FGF21 G198R (SEQ ID NO: 18) 
                 1.15 
               
               
                   
               
               
                 FGF21 G198K (SEQ ID NO: 19) 
                 1.81 
               
               
                   
               
               
                 FGF21 G198Y (SEQ ID NO: 20) 
                 0.68 
               
               
                   
               
               
                 FGF21 P199G (SEQ ID NO: 21) 
                 1.65 
               
               
                   
               
               
                 FGF21 P199T (SEQ ID NO: 23) 
                 1.66 
               
               
                   
               
               
                 FGF21 G198P, P199G (SEQ ID NO: 24) 
                 10.75 
               
               
                   
               
               
                 FGF21 S200Q, Q2015 (SEQ ID NO: 25) 
                 1.62 
               
               
                   
               
               
                 FGF21 H60C, S79C (SEQ ID NO: 75) 
                 67.06 
               
               
                   
               
               
                 FGF21 H145C, L165C (SEQ ID NO: 76) 
                 inactive 
               
               
                   
               
               
                 FGF21 A59C, G71C (SEQ ID NO: 77) 
                 2.15 
               
               
                   
               
               
                 FGF21 H60C, A72C (SEQ ID NO: 78) 
                 inactive 
               
               
                   
               
               
                 FGF21 I63C, G112C (SEQ ID NO: 79) 
                 5.05 
               
               
                   
               
               
                 FGF21 L49C, L170C (SEQ ID NO: 80) 
                 inactive 
               
               
                   
               
               
                 FGF21 T51C, L167C (SEQ ID NO: 81) 
                 inactive 
               
               
                   
               
               
                 FGF21 H140C, L142C (SEQ ID NO: 82) 
                 2.27 
               
               
                   
               
               
                 FGF21 D130C, Y132C (SEQ ID NO: 83) 
                 49.39 
               
               
                   
               
               
                 FGF21 S137C, H140C (SEQ ID NO: 84) 
                 3.87 
               
               
                   
               
               
                 FGF21 S76C, S79C (SEQ ID NO: 85) 
                 1.44 
               
               
                   
               
               
                 FGF21 G108C, L142C (SEQ ID NO: 86) 
                 1.48 
               
               
                   
               
               
                 FGF21 A72C, S79C (SEQ ID NO: 87) 
                 11.85 
               
               
                   
               
               
                 FGF21 H60C, S79C, H145C, L165C (SEQ ID NO: 
                 inactive 
               
               
                 88) 
                   
               
               
                   
               
               
                 FGF21 A72C, S79C, G108C, L142C (SEQ ID NO: 
                 8.24 
               
               
                 89) 
                   
               
               
                   
               
               
                 FGF21 R47C, P174C (SEQ ID NO: 90) 
                 14.53 
               
               
                   
               
               
                 FGF21 M196C, R203C (SEQ ID NO: 91) 
                 inactive 
               
               
                   
               
               
                 FGF21 Q55C, G148C (SEQ ID NO: 93) 
                 0.48 
               
               
                   
               
               
                 FGF21 Q55C, G148C, N149-R159 deleted and 
                 8.19 
               
               
                 replaced by GSGS (SEQ ID NO: 94) 
                   
               
               
                   
               
               
                 FGF21 Q55C, G148C, N149-R159 deleted and 
                 7.27 
               
               
                 replaced by GGSGGS (SEQ ID NO: 95) 
                   
               
               
                   
               
               
                 FGF21 Q55G, Q56C, L146C (SEQ ID NO: 96) 
                 163.20 
               
               
                   
               
               
                 FGF21 Q55G, Q56C, L146C, P147-R159 deleted 
                 113.90 
               
               
                 and replaced by GSGS (SEQ ID NO: 97) 
                   
               
               
                   
               
               
                 FGF21 Q55G, Q56C, L146C, P147-R159 deleted 
                 155.20 
               
               
                 and replaced by GGSGGS (SEQ ID NO: 98) 
                   
               
               
                   
               
               
                 FGF21 R124C, Q136C (SEQ ID NO: 115) 
                 22.59 
               
               
                   
               
               
                 FGF21 P143C, A162C (SEQ ID NO: 116) 
                 17.58 
               
               
                   
               
               
                 FGF21 P77C, K97C (SEQ ID NO: 117) 
                 8.76 
               
               
                   
               
               
                 FGF21 E78C, T98C (SEQ ID NO: 118) 
                 9.22 
               
               
                   
               
               
                 FGF21 L80C, V96C (SEQ ID NO: 119) 
                 17.85 
               
               
                   
               
               
                 FGF21 S195C, S200C (SEQ ID NO: 8) 
                 10.79 
               
               
                   
               
               
                 FGF21 GGGGS inserted between G198 and P199 
                 3.07 
               
               
                 (SEQ ID NO: 15) 
                   
               
               
                   
               
               
                 FGF21 P199R (SEQ ID NO: 22) 
                 0.95 
               
               
                   
               
               
                 FGF21 A59C, A73C (SEQ ID NO: 105) 
                 19.21 
               
               
                   
               
               
                 FGF21 Q56C, L146C (SEQ ID NO: 110) 
                 inactive 
               
               
                   
               
               
                 FGF21 H145C, P161C (SEQ ID NO: 120) 
                 6.51 
               
               
                   
               
               
                 FGF21 E62C, E78C (SEQ ID NO: 101) 
                 2.44 
               
               
                   
               
               
                 FGF21 E62C, S79C (SEQ ID NO: 102) 
                 0.55 
               
               
                   
               
               
                 FGF21 R100C, L114C (SEQ ID NO: 103) 
                 48.22 
               
               
                   
               
               
                 FGF21 L102C, G112C (SEQ ID NO: 104) 
                 inactive 
               
               
                   
               
               
                 FGF21 E62C, A72C (SEQ ID NO: 106) 
                 2.03 
               
               
                   
               
               
                 FGF21 Q104C, 5137C (SEQ ID NO: 107) 
                 1.83 
               
               
                   
               
               
                 FGF21 Q104C, H140C (SEQ ID NO: 108) 
                 21.85 
               
               
                   
               
               
                 FGF21 A54C, P161C (SEQ ID NO: 109) 
                 6.38 
               
               
                   
               
               
                 FGF21 I63C, T98C (SEQ ID NO: 111) 
                 87.58 
               
               
                   
               
               
                 FGF21 G67C, R100C (SEQ ID NO: 112) 
                 23.32 
               
               
                   
               
               
                 FGF21 V69C, L110C (SEQ ID NO: 113) 
                 11.70 
               
               
                   
               
               
                 FGF21 A120C, A139C (SEQ ID NO: 114) 
                 22.00 
               
               
                   
               
               
                 FGF21 L81C, G95C (SEQ ID NO: 121) 
                 46.46 
               
               
                   
               
               
                 FGF21 Q82C, L94C (SEQ ID NO: 122) 
                 5.57 
               
               
                   
               
               
                 FGF21 L83C, I91C (SEQ ID NO: 123) 
                 inactive 
               
               
                   
               
               
                 FGF21 D155L (SEQ ID NO: 124) 
                 0.11 
               
               
                   
               
               
                 FGF21 D155K (SEQ ID NO: 125) 
                 0.06 
               
               
                   
               
               
                 FGF21 D155Y (SEQ ID NO: 126) 
                 0.07 
               
               
                   
               
               
                 FGF21 D155P (SEQ ID NO: 127) 
                 0.08 
               
               
                   
               
               
                 FGF21 D155E (SEQ ID NO: 128) 
                 0.13 
               
               
                   
               
               
                 FGF21 D155N (SEQ ID NO: 129) 
                 0.06 
               
               
                   
               
               
                 FGF21 R154Q, D155L (SEQ ID NO: 130) 
                 0.14 
               
               
                   
               
               
                 FGF21 R154Q, D155K (SEQ ID NO: 131) 
                 0.39 
               
               
                   
               
               
                 FGF21 H153Q, R154Q, D155N, A157V, R159K 
                 0.23 
               
               
                 (SEQ ID NO: 132) 
                   
               
               
                   
               
               
                 FGF21 P152A, H153K, R154K, D155E, P156A, 
                 0.24 
               
               
                 A1575, R159Q (SEQ ID NO: 133) 
                   
               
               
                   
               
               
                 FGF21 H153Y, R154K, D155N, P156K, A157G 
                 0.05 
               
               
                 (SEQ ID NO: 134) 
                   
               
               
                   
               
               
                 FGF21 P152-H153 deleted (SEQ ID NO: 135) 
                 0.21 
               
               
                   
               
               
                 FGF21 N149-R163 deleted and replaced by GSGS 
                 3.69 
               
               
                 (SEQ ID NO: 136) 
                   
               
               
                   
               
               
                 FGF21 N149-A162 deleted and replaced by GSHSG 
                 0.57 
               
               
                 (SEQ ID NO: 137) 
                   
               
               
                   
               
               
                 FGF21 N149-A162 deleted and replaced by 
                 0.19 
               
               
                 GSHSGS (SEQ ID NO: 138) 
                   
               
               
                   
               
               
                 FGF21 K150H (SEQ ID NO: 139) 
                 0.15 
               
               
                   
               
               
                 FGF21 K150H, P152L (SEQ ID NO: 140) 
                 0.13 
               
               
                   
               
               
                 FGF21 R163H (SEQ ID NO: 141) 
                 0.39 
               
               
                   
               
               
                 FGF21 P158H, R159H (SEQ ID NO: 142) 
                 0.84 
               
               
                   
               
               
                 FGF21 N149-D155 deleted and replaced by GSGS 
                 0.28 
               
               
                 (SEQ ID NO: 143) 
                   
               
               
                   
               
               
                 FGF21 N149-D155 deleted and replaced by GSHSG 
                 0.10 
               
               
                 (SEQ ID NO: 144) 
                   
               
               
                   
               
               
                 FGF21 N149-D155 deleted and replaced by ATTS 
                 0.02 
               
               
                 (SEQ ID NO: 145) 
                   
               
               
                   
               
               
                 FGF21 R159-R163 deleted and replaced by HE 
                 0.98 
               
               
                 (SEQ ID NO: 150) 
                   
               
               
                   
               
               
                 FGF21 A162Y (SEQ ID NO: 151) 
                 0.08 
               
               
                   
               
               
                 FGF21 R159H (SEQ ID NO: 152) 
                 0.17 
               
               
                   
               
               
                 FGF21 S200M (SEQ ID NO: 153) 
                 0.17 
               
               
                   
               
               
                 FGF21 G202T (SEQ ID NO: 154) 
                 0.01 
               
               
                   
               
               
                 FGF21 G202-R203 deleted (SEQ ID NO: 157) 
                 1.63 
               
               
                   
               
               
                 FGF21 G198 deleted (SEQ ID NO: 29) 
                 41.65 
               
               
                   
               
               
                 FGF21 V197 deleted (SEQ ID NO: 34) 
                 68.8 
               
               
                   
               
               
                 FGF21 S76C, S79C, G198Y (SEQ ID NO: 39) 
                 0.15 
               
               
                   
               
               
                 FGF21 Q55C, P147C, P199 deleted (SEQ ID NO: 
                 10.03 
               
               
                 44) 
                   
               
               
                   
               
               
                 FGF21 G198-P199 deleted (SEQ ID NO: 30) 
                 14.47 
               
               
                   
               
               
                 FGF21 G198 deleted, P199F (SEQ ID NO: 31) 
                 2.10 
               
               
                   
               
               
                 FGF21 G198 deleted, P199L (SEQ ID NO: 32) 
                 4.80 
               
               
                   
               
               
                 FGF21 G198 deleted, P199L, S200N (SEQ ID NO: 
                 9.44 
               
               
                 33) 
                   
               
               
                   
               
               
                 FGF21 L146C, A162C (SEQ ID NO: 35) 
                 8.28 
               
               
                   
               
               
                 FGF21 A59C, G71C, P199 deleted (SEQ ID NO: 36) 
                 1.16 
               
               
                   
               
               
                 FGF21 A59C, G71C, G198Y (SEQ ID NO: 37) 
                 0.75 
               
               
                   
               
               
                 FGF21 S76C, S79C, P199 deleted (SEQ ID NO: 38) 
                 1.25 
               
               
                   
               
               
                 FGF21 G108C, L142C, P199 deleted (SEQ ID NO: 
                 0.90 
               
               
                 40) 
                   
               
               
                   
               
               
                 FGF21 G108C, L142C, G198Y (SEQ ID NO: 41) 
                 0.55 
               
               
                   
               
               
                 FGF21 Q55C, P147C, G198Y (SEQ ID NO: 45) 
                 3.60 
               
               
                   
               
               
                 FGF21 Q55C, N149C, P199 deleted (SEQ ID NO: 
                 1.70 
               
               
                 46) 
                   
               
               
                   
               
               
                 FGF21 Q55C, N149C, G198Y (SEQ ID NO: 47) 
                 0.58 
               
               
                   
               
               
                 FGF21 PLSMVGPSQGRSPSYAS inserted after 
                 11.26 
               
               
                 S209 (SEQ ID NO: 159) 
                   
               
               
                   
               
               
                 FGF21 P199 deleted, PLSMVGSQGRSPSYAS 
                 15.73 
               
               
                 inserted after S209 (SEQ ID NO: 160) 
                   
               
               
                   
               
               
                 FGF21 V197-G198 deleted (SEQ ID NO: 72) 
                 inactive 
               
               
                   
               
               
                 FGF21 M196 deleted (SEQ ID NO: 48) 
                 inactive 
               
               
                   
               
               
                 FGF21 S190-S191 deleted (SEQ ID NO: 54) 
                 inactive 
               
               
                   
               
               
                 FGF21 Q184 deleted (SEQ ID NO: 60) 
                 2.26 
               
               
                   
               
               
                 FGF21 P205 deleted (SEQ ID NO: 66) 
                 27.38 
               
               
                   
               
               
                 FGF21 S195 deleted (SEQ ID NO: 49) 
                 inactive 
               
               
                   
               
               
                 FGF21 L194 deleted (SEQ ID NO: 50) 
                 inactive 
               
               
                   
               
               
                 FGF21 P193 deleted (SEQ ID NO: 51) 
                 inactive 
               
               
                   
               
               
                 FGF21 D192 deleted (SEQ ID NO: 52) 
                 inactive 
               
               
                   
               
               
                 FGF21 S191 deleted (SEQ ID NO: 53) 
                 97.28 
               
               
                   
               
               
                 FGF21 G189 deleted (SEQ ID NO: 55) 
                 76.16 
               
               
                   
               
               
                 FGF21 V188 deleted (SEQ ID NO: 56) 
                 inactive 
               
               
                   
               
               
                 FGF21 D187 deleted (SEQ ID NO: 57) 
                 86.1 
               
               
                   
               
               
                 FGF21 P186 deleted (SEQ ID NO: 58) 
                 0.87 
               
               
                   
               
               
                 FGF21 P185-P186 deleted (SEQ ID NO: 59) 
                 21.09 
               
               
                   
               
               
                 FGF21 S200 deleted (SEQ ID NO: 61) 
                 1.17 
               
               
                   
               
               
                 FGF21 Q201 deleted (SEQ ID NO: 62) 
                 1.91 
               
               
                   
               
               
                 FGF21 G202 deleted (SEQ ID NO: 63) 
                 8.18 
               
               
                   
               
               
                 FGF21 R203 deleted (SEQ ID NO: 64) 
                 6.69 
               
               
                   
               
               
                 FGF21 S204 deleted (SEQ ID NO: 65) 
                 25.67 
               
               
                   
               
               
                 FGF21 S206 deleted (SEQ ID NO: 67) 
                 31.89 
               
               
                   
               
               
                 FGF21 Y207 deleted (SEQ ID NO: 68) 
                 7.14 
               
               
                   
               
               
                 FGF21 V197 deleted, S201 deleted (SEQ ID NO: 69) 
                 98.7 
               
               
                   
               
               
                 FGF21 M196 deleted, S201 deleted (SEQ ID NO: 
                 165.3 
               
               
                 70) 
                   
               
               
                   
               
               
                 FGF21 M196-V197 deleted (SEQ ID NO: 71) 
                 inactive 
               
               
                   
               
               
                 FGF21 S195-M196 deleted (SEQ ID NO: 73) 
                 inactive 
               
               
                   
               
               
                 FGF21 G198 deleted, S200 deleted (SEQ ID NO: 
                 77.77 
               
               
                 74) 
               
               
                   
               
            
           
         
       
     
     Example 4: In Vitro Activity Measurements of FGF21 Variants with Primary Hman Adipocytes 
     The cellular in vitro efficacy of human FGF21 (SEQ ID NO: 2) or FGF21 variants was also measured with human primary subcutaneous adipocytes using In-Cell Western (ICW) assay (Aguilar H. N. et al. (2010) PLoS ONE 5(4): e9965). Briefly, 2.8×10 4  human preadipocytes (PromoCell, Heidelberg, Germany) were seeded into each well of 96-well plates and differentiated to mature adipocytes (Hemmrich K. et al. (2005) Differentiation 73(1): 28-35; Lee M. J. (2014) Methods Enzymol. 538: 49-65). Prior to stimulation of the MAP kinase ERK1/2 activation, cells were serum starved with serum-free medium (DMEM 1 g/l /Ham&#39;s F-10 Medium (1:1, v/v) (PAN-Biotech, Aidenbach, Germany), 15 mmol/L Hepes, pH 7.4, 33 μmol/L biotin, 17 pmol/L pantothenate) for 3-4 h. The cells were subsequently treated with increasing concentrations of either mature human FGF21 (SEQ ID NO: 2), the indicated FGF21 variant, or other peptides for 5 min at 37° C. After incubation, the medium was discarded, and the cells were fixed in 3.7% freshly prepared para-formaldehyde for 20 min. Cells were permeabilized with 0.1% Triton-X-100 in PBS for 20 min. Blocking was performed with Odyssey blocking buffer (LICOR, Bad Homburg, Germany) for 2 h at room temperature. As primary antibody, anti-pERK Phospho-p44/42 MAP Kinase Thr202/Tyr204 (Cell Signaling) was added and incubated overnight at 4° C. After incubation of the primary antibody, cells were washed with PBS plus 0.1% Tween20. The secondary anti-Mouse 800CW antibody (LICOR, Bad Homburg, Germany) was incubated for 1 h at room temperature. Subsequently, cells were washed again with PBS plus 0.1% Tween20, and infrared dye signals were quantified with an Odyssey imager (LICOR, Bad Homburg, Germany). Results were normalized by quantification of DNA with TO-PRO3 dye (Invitrogen, Karlsruhe, Germany). Data were obtained as arbitrary units (AU), and EC50 values were obtained from dose-response curves as shown in  FIGS. 4A and 4B . EC50 values are summarized in Table 5. 
     
       
         
           
               
             
               
                 TABLE 5 
               
             
            
               
                   
               
               
                 EC50-values of mature human FGF21 (SEQ ID NO: 2) and FGF21  
               
               
                 variants measured via ICW pERK in primary human adipocytes.  
               
            
           
           
               
               
            
               
                   
                 pERK ICW 
               
               
                 Protein/Mutation 
                 EC50 (nmol/L) 
               
               
                   
               
            
           
           
               
               
            
               
                 FGF21, human (SEQ ID NO: 2) 
                 1.55 
               
               
                 FGF21 P199 deleted (SEQ ID NO: 11) 
                 3.47 
               
               
                 FGF21 Q55C, P147C, P199 deleted  
                 5.32 
               
               
                 (SEQ ID NO: 44) 
                   
               
               
                 FGF21 Q55C, N149C, P199 deleted  
                 1.42 
               
               
                 (SEQ ID NO: 46) 
                   
               
               
                 FGF21 G198Y (SEQ ID NO: 20) 
                 1.03 
               
               
                 FGF21 Q55C, P147C, G198Y (SEQ ID NO: 45) 
                 2.55 
               
               
                 FGF21 Q55C, N149C, G198Y (SEQ ID NO: 47) 
                 1.18 
               
               
                   
               
               
                 The numbering of the amino acids refers to human wild-type FGF21 of SEQ ID NO: 1. 
               
            
           
         
       
     
     Example 5: Analyzing Thermal Stability of Human FGF21 and FGF21 Variants 
     To analyze the impact of mutations in FGF21 on thermostability, a thermal shift assay was applied in imitation of differential scanning fluorimetry (DSF or ThermoFluorTM) assays used for searching stabilizing ligands (Ahmad S. et al. (2012) Protein Science 21: 433-446; Pantoliano et al. (2001) J. Biomol. Screen 6: 429-440; Niesen et al. (2007) Nat. Protoc. 2: 2212-21). This assay is based on the observation that hydrophobic fluorescent dyes, such as Sypro™ Orange (Life Technologies, cat. No. S6651), increase their fluorescence when they bind to hydrophobic patches on a protein. Such hydrophobic patches are exposed in proteins when they unfold upon heating, so that the increase in fluorescence can be used as a measure for the degree of unfolding and, hence, for the thermostability of the proteins. 
     FGF21 variants were tested by mixing a solution of the protein in PBS (Gibco) with a 160×solution of SyproT™Orange (diluted in water from a 5000×DMSO stock as provided by the supplier). The sample volume was adjusted to 20 μl with PBS. Typical conditions contained 0.8 mg/ml FGF21 variant protein and 8×Sypro™Orange in the final mixture, but protein concentrations could be varied between 0.4 mg/ml and 1.2 mg/ml. Samples were dispensed in 96-well PCR plates (BioRad Semi-Skirt 96 white) and shortly centrifuged to remove air bubbles. Plates were inserted in a BioRad iQ5 real-time PCR instrument and subjected to a thermal gradient from 10 to 90° C. at a ramp speed of 1° C./min. For excitation and quantification of fluorescence, filters for wavelengths of 485 nm and 575 nm were chosen. Biorad iQ5 Standard Edition software (v. 2.0.148.60623) was used for data processing. In curves of fluorescence intensity against temperature, the inflection point was chosen as the measure for the melting temperature (T M ). Human FGF21 (SEQ ID NO: 3) was included on every plate as a reference.  FIG. 5  is showing representative melting curves of selected FGF21 variants and Table 6 is listing the differences in T M  values with respect to human FGF21 (SEQ ID NO: 3). 
     
       
         
           
               
             
               
                 TABLE 6  
               
             
            
               
                   
               
               
                 Melting point differences (deltaTm) of FGF21 variants with 
               
               
                 respect to human FGF21 of SEQ ID NO: 3. 
               
            
           
           
               
               
               
            
               
                   
                 SEQ ID 
                 deltaTm 
               
               
                 Protein/Mutation 
                 NO 
                 (° C.) 
               
               
                   
               
            
           
           
               
               
               
            
               
                 FGF21, human 
                 3 
                 0 
               
               
                   
               
               
                 FGF21 D155L 
                 124 
                 0 
               
               
                   
               
               
                 FGF21 D155K 
                 125 
                 −1.3 
               
               
                   
               
               
                 FGF21 D155Y 
                 126 
                 −0.7 
               
               
                   
               
               
                 FGF21 D155P 
                 127 
                 0 
               
               
                   
               
               
                 FGF21 D155E 
                 128 
                 0 
               
               
                   
               
               
                 FGF21 D155N 
                 129 
                 −1 
               
               
                   
               
               
                 FGF21 R154Q, D155L 
                 130 
                 −1 
               
               
                   
               
               
                 FGF21 R154Q, D155K 
                 131 
                 −1.7 
               
               
                   
               
               
                 FGF21 H153Q, R154Q, D155N, A157V, R159K 
                 132 
                 −2.3 
               
               
                   
               
               
                 FGF21 P152A, H153K, R154K, D155E, P156A, 
                 133 
                 −2.7 
               
               
                 A1575, R159Q 
                   
                   
               
               
                   
               
               
                 FGF21 H153Y, R154K, D155N, P156K, A157G 
                 134 
                 2.3 
               
               
                   
               
               
                 FGF21 P152-H153 deleted 
                 135 
                 0.3 
               
               
                   
               
               
                 FGF21 N149-R163 deleted and replaced by GSGS 
                 136 
                 −3 
               
               
                 (SEQ ID NO: 161) 
                   
                   
               
               
                   
               
               
                 FGF21 N149-A162 deleted and replaced by GSHSG 
                 137 
                 9.4 
               
               
                 (SEQ ID NO: 163) 
                   
                   
               
               
                   
               
               
                 FGF21 N149-A162 deleted and replaced by 
                 138 
                 2.3 
               
               
                 GSHSGS (SEQ ID NO: 165) 
                   
                   
               
               
                   
               
               
                 FGF21 K150H 
                 139 
                 0.7 
               
               
                   
               
               
                 FGF21 K150H, P152L 
                 140 
                 0.7 
               
               
                   
               
               
                 FGF21 R163H 
                 141 
                 −10.1 
               
               
                   
               
               
                 FGF21 P158H, R159H 
                 142 
                 0.7 
               
               
                   
               
               
                 FGF21 N149-D155 deleted and replaced by GSGS 
                 143 
                 2.9 
               
               
                 (SEQ ID NO: 161) 
                   
                   
               
               
                   
               
               
                 FGF21 N149-D155 deleted and replaced by GSHSG 
                 144 
                 3.6 
               
               
                 (SEQ ID NO: 163) 
                   
                   
               
               
                   
               
               
                 FGF21 N149-D155 deleted and replaced by ATTS 
                 145 
                 2.6 
               
               
                 (SEQ ID NO: 164) 
                   
                   
               
               
                   
               
               
                 FGF21 R159-R163 deleted and replaced by GA 
                 146 
                 −13 
               
               
                   
               
               
                 FGF21 R159-R163 deleted and replaced by GY 
                 147 
                 −17.5 
               
               
                   
               
               
                 FGF21 R159-R163 deleted and replaced by HH 
                 148 
                 12.5 
               
               
                   
               
               
                 FGF21 R159-R163 deleted and replaced by GE 
                 149 
                 −14.5 
               
               
                   
               
               
                 FGF21 R159-R163 deleted and replaced by HE 
                 150 
                 −16.4 
               
               
                   
               
               
                 FGF21 A162Y 
                 151 
                 1 
               
               
                   
               
               
                 FGF21 R159H 
                 152 
                 0 
               
               
                   
               
               
                 FGF21 S200M 
                 153 
                 0.7 
               
               
                   
               
               
                 FGF21 G202T 
                 154 
                 0.3 
               
               
                   
               
               
                 FGF21 R203E 
                 155 
                 −1.5 
               
               
                   
               
               
                 FGF21 R203H 
                 156 
                 −1 
               
               
                   
               
               
                 FGF21 G202-R203 deleted 
                 157 
                 1.7 
               
               
                   
               
               
                 FGF21 G108C, L142C 
                 86 
                 [14] 
               
               
                   
               
               
                 FGF21 M196C, R203C 
                 91 
                 [−1] 
               
               
                   
               
               
                 FGF21 S76C, S79C 
                 85 
                 8 
               
               
                   
               
               
                 FGF21 H140C, L142C 
                 82 
                 [−10] 
               
               
                   
               
               
                 FGF21 Q55C, P147C 
                 99 
                 20 
               
               
                   
               
               
                 FGF21 Q55C, N149C 
                 100 
                 22 
               
               
                   
               
               
                 FGF21 R47C, P174C 
                 90 
                 7.5 
               
               
                   
               
               
                 FGF21 A59C, G71C 
                 77 
                 23.5 
               
               
                   
               
               
                 FGF21 H60C, S79C 
                 75 
                 19.9 
               
               
                   
               
               
                 FGF21 H145C, L165C 
                 76 
                 −8.5 
               
               
                   
               
               
                 FGF21 H60C, A72C 
                 78 
                 n.d. 
               
               
                   
               
               
                 FGF21 I63C, G112C 
                 79 
                 n.d. 
               
               
                   
               
               
                 FGF21 L49C, L170C 
                 80 
                 n.d. 
               
               
                   
               
               
                 FGF21 T51C, L167C 
                 81 
                 n.d. 
               
               
                   
               
               
                 FGF21 D130C, Y132C 
                 83 
                 n.d. 
               
               
                   
               
               
                 FGF21 S137C, H140C 
                 84 
                 n.d. 
               
               
                   
               
               
                 FGF21 A72C, S79C 
                 87 
                 n.d. 
               
               
                   
               
               
                 FGF21 H60C, S79C, H145C, L165C 
                 88 
                 n.d. 
               
               
                   
               
               
                 FGF21 A72C, S79C, G108C, L142C 
                 89 
                 n.d. 
               
               
                   
               
               
                 FGF21 M196C, R203C 
                 91 
                 n.d. 
               
               
                   
               
               
                 FGF21 Q55C, G148C, N149-R159 deleted and 
                 94 
                 n.d. 
               
               
                 replaced by GSGS (SEQ ID NO: 161) 
                   
                   
               
               
                   
               
               
                 FGF21 Q55C, N149C, G198Y 
                 47 
                 21.9 
               
               
                   
               
               
                 FGF21 Q55C, N149C, P199 deleted 
                 46 
                 22.2 
               
               
                   
               
               
                 FGF21 Q55C, P147C, G198Y 
                 45 
                 20.7 
               
               
                   
               
               
                 FGF21 Q55C, P147C, P199 deleted 
                 44 
                 21.1 
               
               
                   
               
               
                 FGF21 Q55C, G148C, N149-R159 deleted and 
                 95 
                 n.d. 
               
               
                 replaced by GGSGGS (SEQ ID NO: 162) 
                   
                   
               
               
                   
               
               
                 FGF21 Q55G, Q56C, L146C 
                 96 
                 n.d. 
               
               
                   
               
               
                 FGF21 Q55G, Q56C, L146C, P147-R159 deleted 
                 97 
                 n.d. 
               
               
                 and replaced by GSGS (SEQ ID NO: 161) 
                   
                   
               
               
                   
               
               
                 FGF21 Q55G, Q56C, L146C, P147-R159 deleted 
                 98 
                 n.d. 
               
               
                 and replaced by GGSGGS (SEQ ID NO: 162) 
                   
                   
               
               
                   
               
               
                 FGF21 R124C, Q136C 
                 115 
                 n.d. 
               
               
                   
               
               
                 FGF21 P143C, A162C 
                 116 
                 n.d. 
               
               
                   
               
               
                 FGF21 P77C, K97C 
                 117 
                 n.d. 
               
               
                   
               
               
                 FGF21 L80C, V96C 
                 119 
                 n.d. 
               
               
                   
               
               
                 FGF21 S195C, S200C 
                 8 
                 n.d. 
               
               
                   
               
               
                 FGF21 GGGGS (SEQ ID NO: 168) inserted between 
                 15 
                 n.d. 
               
               
                 G198 and P199 
                   
                   
               
               
                   
               
               
                 FGF21 A59C, A73C 
                 105 
                 n.d. 
               
               
                   
               
               
                 FGF21 Q56C, L146C 
                 110 
                 n.d. 
               
               
                   
               
               
                 Values in brackets represent data from curves with initial high fluorescence and are less reliable. 
               
               
                 “n.d.”: not determinable Tested variants may carry an N-terminal His-tag (SEQ ID NO: 5 or 6). 
               
               
                 The tag may also be removed by protease-cleavage. 
               
               
                 The numbering of the amino acids refers to human wild-type FGF21 of SEQ ID NO: 1. 
               
            
           
         
       
     
     Example 6: Design of Thermostable Variants and X-ray Crystal Structure of FGF21 Q55C, N149C (SEQ ID NO: 158) 
     Since in public data bases like RCSB Protein Data Bank (PDB) no structure of FGF21 was deposited, different homology models of human FGF21 (SEQ ID NO: 2) were created using the SWISS-MODEL homology modeling server (Kiefer et al. (2009) Nucleic Acids Res. 37: D387-D392) or the software program BioLuminate (BioLuminate, version 1.0, Schrodinger, LLC, New York, N.Y., 2012). These models are based on the published crystal structures of FGF19 (Goetz et al. (2007) Mol. Cell. Biol. 27: 3417-3428; PDB code 2P23; Harmer (2004) Biochemistry 43: 629-640; PDB code 1PWA) and FGF23 (Goetz et al. (2007) Mol. Cell. Biol. 27: 3417-3428; PDB code 2P39) ( FIGS. 6A-6C ). The different models were subjected to further analysis using BioLuminate to generate a list of potential sites for introducing disulfide bridges. Potential positions were inspected visually in the model structures using the software program PYMOL (The PyMOL Molecular Graphics System, Version 1.7 Schrodinger, LLC) for the final selection of candidates for expression and experimental testing. 
     To crystallize human FGF21, a modified and truncated variant (human FGF21 S34-E176, Q55C, N149C; SEQ ID NO: 158) was expressed and purified as described above (Example 1). 
     Crystals were grown by hanging-drop vapor diffusion. The protein was concentrated to 12 mg/mL in 50 mmol/L Tris, pH 8.0, 0.5 mol/L NaCl. The reservoir solution contained 2 mol/L NH 4 SO 4  in 100 mmol/L Tris, pH 8.5. 100 nL protein solution was mixed with 100 nL reservoir solution and equilibrated at 20° C. Crystals appeared within two weeks. Cryoprotection was achieved by soaking crystals in the reservoir solution supplemented with 20 to 25% (vol/vol) ethylene glycol before flash freezing in liquid nitrogen. 
     X-ray diffraction data were collected at beamline PX-III of the Swiss Light Source (SLS) in Viligen, Switzerland, and processed with XDS (Kabsch W. (2010) Acta Crystallogr. D66: 125-132) and scaled with Aimless (Evans P. R. (2006) Acta Crystallogr. D62: 72-82) as implemented in autoProc (Vonrhein C. et al. (2011) Acta Crystallogr. D67: 293-302). The crystal was of space group P4 1 32 and contained two FGF21 Q55C, N149C molecules in the asymmetric unit. The unit cell dimensions of this crystal were as follows: a=b=c=136.7 A. The crystal diffracted to 2.97 Å resolution and the Rmerge was 10.5%. 
     The structure was determined by molecular replacement using the program Phaser (McCoy A. J. et al. (2007) J. Appl. Crystallogr. 40: 658-674). The crystal structures of FGF19 (Protein Data Bank code 1pwa), FGF23 (Protein Data Bank code 2p39), and FGF9 (Protein Data Bank code 2P39) were superimposed, and all loops that differed more than ˜5 A were removed. The resulting ensemble was used as a search model and resulted in an unambiguous solution. 
     Phaser placed three FGF-ensembles. However, the electron density for the third ensemble was extremely weak, and this ensemble was deleted. Model building was done with Coot (Emsley P. et al. (2010) Acta Crystallogr. D66: 486-501) and refinement was done with Buster (Bricogne G. et al. (2011) Cambridge, United Kingdom: Global Phasing Ltd.) and Refmac5 (Murshudov G.N. et al. (2011) Acta Crystallogr. D67: 355-367). The final Rfactor was 25.5% and the free Rfactor was 28.6%. These Rfactors were relatively high and may indicate a certain amount of disorder in the crystals. 
     In the final model for the FGF21 Q55C, N149C (SEQ ID NO: 158), molecule A contained the residues G42-K150 and P158-G169; molecule B contained the residues P36-H154 and P156-L170. In both molecules, the loop between P150 and R159 was significantly disordered which is an indication for high flexibility in this region. 
     It may be assumed that such a highly flexible loop represents a starting point for thermal unfolding of the protein. Hence, stabilization of this region by a disulfide bond may explain the significant increase in thermal stability of about 20° C. as assessed for the FGF21 variant H29-S209 Q55C, N149C (SEQ ID NO: 100) or the neighboring variant H29-5209 Q55C, P147C (SEQ ID NO: 99). 
     
       
         
           
               
             
               
                 TABLE 7 
               
             
            
               
                   
               
               
                 List of Sequences. 
               
            
           
           
               
               
               
            
               
                 SEQ 
                   
                   
               
               
                 ID NO 
                 Description 
                 Sequence 
               
               
                   
               
            
           
           
               
               
               
            
               
                 1 
                 full-length human 
                 MDSDETGFEH SGLWVSVLAG LLLGACQAHP IPDSSPLLQF GGQVRQRYLY 
               
               
                   
                 wild-type FGF21 
                 TDDAQQTEAH LEIREDGTVG GAADQSPESL LQLKALKPGV IQILGVKTSR 
               
               
                   
                 (including signal 
                 FLCQRPDGAL YGSLHFDPEA CSFRELLLED GYNVYQSEAH GLPLHLPGNK 
               
               
                   
                 sequence Metl- 
                 SPHRDPAPRG PARFLPLPGL PPAPPEPPGI LAPQPPDVGS SDPLSMVGPS 
               
               
                   
                 Ala28) 
                 QGRSPSYAS 
               
               
                   
               
               
                 2 
                 mature human 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 wild-type FGF21 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 (without signal 
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                 sequence Metl- 
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
                 Ala28) 
                   
               
               
                   
               
               
                 3 
                 mature human 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 wild-type FGF21 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 with additional N- 
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                 terminal G (G- 
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
                 FGF21) 
                   
               
               
                   
               
               
                 4 
                 IL2 signal 
                 MYRMQLLSCI ALLSLALVTN S 
               
               
                   
                 sequence 
                   
               
               
                   
               
               
                 5 
                 Histidine-rich 
                 DIQTGGSHAH GHGHAHGHGG SGENLYFQ 
               
               
                   
                 presequence a 
                   
               
               
                   
                 (His-tag + TEV 
                   
               
               
                   
                 cleavage site) 
                   
               
               
                   
               
               
                 6 
                 Histidine-rich 
                 MGHHHHHHHH GGGENLYFQG 
               
               
                   
                 presequence b 
                   
               
               
                   
                 (His-tag + TEV 
                   
               
               
                   
                 cleavage site) 
                   
               
               
                   
               
               
                 7 
                 Histidine-rich 
                 MGHHHHHHGS LQDSEVNQEA KPEVKPEVKP ETHINLKVSD GSSEIFFKIK 
               
               
                   
                 presequence c 
                 KTTPLRRLME AFAKRQGKEM DSLRFLYDGI RIQADQAPED LDMEDNDIIE 
               
               
                   
                 (His-tag) + 
                 AHREQIGG 
               
               
                   
                 SUMO-tag 
                   
               
               
                   
               
               
                 8 
                 G-FGF21 S195C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 S200C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLCMV GPCQGRSPSY AS 
               
               
                   
               
               
                 9 
                 G-FGF21 P199S, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 S200P 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GSPQGRSPSY AS 
               
               
                   
               
               
                 10 
                 G-FGF21 P199Q, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 Q201P 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GQSPGRSPSY AS 
               
               
                   
               
               
                 11 
                 G-FGF21 P199 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GSQGRSPSYA S 
               
               
                   
               
               
                 12 
                 G-FGF21 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 M196P,V197G, 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 G198V,P199M 
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSPG VMSQGRSPSY AS 
               
               
                   
               
               
                 13 
                 G-FGF21 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 M196V,V197M 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSVM GPSQGRSPSY AS 
               
               
                   
               
               
                 14 
                 G-FGF21 V197E 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                   
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSME GPSQGRSPSY AS 
               
               
                   
               
               
                 15 
                 G-FGF21 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 GGGGS (SEQ ID 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 NO: 168) inserted 
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                 between G198 
                 PGILAPQPPD VGSSDPLSMV GGGGGSPSQG RSPSYAS 
               
               
                   
                 and P199 
                   
               
               
                   
               
               
                 16 
                 G-FGF21 G198E 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                   
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV EPSQGRSPSY AS 
               
               
                   
               
               
                 17 
                 G-FGF21 G1980 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                   
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV DPSQGRSPSY AS 
               
               
                   
               
               
                 18 
                 G-FGF21 G198R 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                   
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV RPSQGRSPSY AS 
               
               
                   
               
               
                 19 
                 G-FGF21 G198K 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                   
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV KPSQGRSPSY AS 
               
               
                   
               
               
                 20 
                 G-FGF21 G198Y 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                   
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV YPSQGRSPSY AS 
               
               
                   
               
               
                 21 
                 G-FGF21 P199G 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                   
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GGSQGRSPSY AS 
               
               
                   
               
               
                 22 
                 G-FGF21 P199R 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                   
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GRSQGRSPSY AS 
               
               
                   
               
               
                 23 
                 G-FGF21 P199T 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                   
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GTSQGRSPSY AS 
               
               
                   
               
               
                 24 
                 G-FGF21 G198P, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 P199G 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV PGSQGRSPSY AS 
               
               
                   
               
               
                 25 
                 G-FGF21 5200Q, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 Q201S 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPQSGRSPSY AS 
               
               
                   
               
               
                 26 
                 G-FGF21 V197C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 G198V, P199C, 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 S200H 
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMD VDHQGRSPSY AS 
               
               
                   
               
               
                 27 
                 G-FGF21 V197- 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 R203 deleted and 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 replaced by 
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                 GHRSHLQTVF 
                 PGILAPQPPD VGSSDPLSMG HRSHLQTVFS PSYAS 
               
               
                   
                 (SEQ ID NO: 166) 
                   
               
               
                   
               
               
                 28 
                 G-FGF21 V197- 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 R203 deleted and 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 replaced by 
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                 GLNSMV (SEQ 
                 PGILAPQPPD VGSSDPLSMG LNSMVSPSYA S 
               
               
                   
                 ID NO: 167) 
                   
               
               
                   
               
               
                 29 
                 G-FGF21 G198 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV PSQGRSPSYA S 
               
               
                   
               
               
                 30 
                 G-FGF21 G198- 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 P199 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV SQGRSPSYAS 
               
               
                   
               
               
                 31 
                 G-FGF21 G198 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted, P199F 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV FSQGRSPSYA S 
               
               
                   
               
               
                 32 
                 G-FGF21 G198 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted, P199L 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV LSQGRSPSYA S 
               
               
                   
               
               
                 33 
                 G-FGF21 G198 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted, P199L, 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 S200N 
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV LNQGRSPSYA S 
               
               
                   
               
               
                 34 
                 G-FGF21 V197 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMG PSQGRSPSYA S 
               
               
                   
               
               
                 35 
                 G-FGF21 L146C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 A162C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHCP GNKSPHRDPA PRGPCRFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 36 
                 G-FGF21 A59C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT ECHLEIREDG TVGCAADQSP 
               
               
                   
                 G71C, P199 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 deleted 
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GSQGRSPSYA S 
               
               
                   
               
               
                 37 
                 G-FGF21 A59C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT ECHLEIREDG TVGCAADQSP 
               
               
                   
                 G71C, G198Y 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV YPSQGRSPSY AS 
               
               
                   
               
               
                 38 
                 G-FGF21 S76C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQCP 
               
               
                   
                 S79C, P199 
                 ECLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 deleted 
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GSQGRSPSYA S 
               
               
                   
               
               
                 39 
                 G-FGF21 S76C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQCP 
               
               
                   
                 S79C, G198Y 
                 ECLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV YPSQGRSPSY AS 
               
               
                   
               
               
                 40 
                 G-FGF21 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 G108C, L142C, 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD CALYGSLHFD PEACSFRELL 
               
               
                   
                 P199 deleted 
                 LEDGYNVYQS EAHGCPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GSQGRSPSYA S 
               
               
                   
               
               
                 41 
                 G-FGF21 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 G108C, L142C, 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD CALYGSLHFD PEACSFRELL 
               
               
                   
                 G198Y 
                 LEDGYNVYQS EAHGCPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV YPSQGRSPSY AS 
               
               
                   
               
               
                 42 
                 G-FGF21 D107C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 D155C, P199 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPC GALYGSLHFD PEACSFRELL 
               
               
                   
                 deleted 
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRCPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GSQGRSPSYA S 
               
               
                   
               
               
                 43 
                 G-FGF21 D107C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 D155C, G198Y 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPC GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRCPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV YPSQGRSPSY AS 
               
               
                   
               
               
                 44 
                 G-FGF21 Q55C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDACQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 P1470, P199 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 deleted 
                 LEDGYNVYQS EAHGLPLHLC GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GSQGRSPSYA S 
               
               
                   
               
               
                 45 
                 G-FGF21 Q55C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDACQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 P147C, G198Y 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLC GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV YPSQGRSPSY AS 
               
               
                   
               
               
                 46 
                 G-FGF21 Q55C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDACQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 N149C, P199 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 deleted 
                 LEDGYNVYQS EAHGLPLHLP GCKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GSQGRSPSYA S 
               
               
                   
               
               
                 47 
                 G-FGF21 Q55C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDACQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 N149C, G198Y 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GCKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV YPSQGRSPSY AS 
               
               
                   
               
               
                 48 
                 G-FGF21 M196 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSVG PSQGRSPSYA S 
               
               
                   
               
               
                 49 
                 G-FGF21 S195 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLMVG PSQGRSPSYA S 
               
               
                   
               
               
                 50 
                 G-FGF21 L194 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 51 
                 G-FGF21 P193 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 52 
                 G-FGF21 D192 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 53 
                 G-FGF21 S191 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 54 
                 G-FGF21 S190- 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 S191 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGDPLSMVGP SQGRSPSYAS 
               
               
                   
               
               
                 55 
                 G-FGF21 G189 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 56 
                 G-FGF21 V188 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 57 
                 G-FGF21 D187 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 58 
                 G-FGF21 P186 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 59 
                 G-FGF21 P185- 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 P186 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQDVG SSDPLSMVGP SQGRSPSYAS 
               
               
                   
               
               
                 60 
                 G-FGF21 Q184 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 61 
                 G-FGF21 S200 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPQGRSPSYA S 
               
               
                   
               
               
                 62 
                 G-FGF21 Q201 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSGRSPSYA S 
               
               
                   
               
               
                 63 
                 G-FGF21 G202 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQRSPSYA S 
               
               
                   
               
               
                 64 
                 G-FGF21 R203 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGSPSYA S 
               
               
                   
               
               
                 65 
                 G-FGF21 S204 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRPSYA S 
               
               
                   
               
               
                 66 
                 G-FGF21 P205 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSSYA S 
               
               
                   
               
               
                 67 
                 G-FGF21 S206 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPYA S 
               
               
                   
               
               
                 68 
                 G-FGF21 Y207 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSA S 
               
               
                   
               
               
                 69 
                 G-FGF21 V197 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted, S201 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 deleted 
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMG PQGRSPSYAS 
               
               
                   
               
               
                 70 
                 G-FGF21 M196 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted, S201 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 deleted 
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSVG PQGRSPSYAS 
               
               
                   
               
               
                 71 
                 G-FGF21 M196- 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 V197 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSGP SQGRSPSYAS 
               
               
                   
               
               
                 72 
                 G-FGF21 V197- 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 G198 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMP SQGRSPSYAS 
               
               
                   
               
               
                 73 
                 G-FGF21 S195- 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 M196 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLVGP SQGRSPSYAS 
               
               
                   
               
               
                 74 
                 G-FGF21 G198 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted, S200 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 deleted 
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV PQGRSPSYAS 
               
               
                   
               
               
                 75 
                 G-FGF21 H60C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EACLEIREDG TVGGAADQSP 
               
               
                   
                 S79C 
                 ECLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 76 
                 G-FGF21 H145C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 L165C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLCLP GNKSPHRDPA PRGPARFCPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 77 
                 G-FGF21 A59C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT ECHLEIREDG TVGCAADQSP 
               
               
                   
                 G71C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 78 
                 G-FGF21 H60C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EACLEIREDG TVGGCADQSP 
               
               
                   
                 A72C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 79 
                 G-FGF21 I63C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLECREDG TVGGAADQSP 
               
               
                   
                 G112C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYCSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 80 
                 G-FGF21 L49C, 
                 GHPIPDSSPL LQFGGQVRQR YCYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 L170C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGCPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 81 
                 G-FGF21 T51C, 
                 GHPIPDSSPL LQFGGQVRQR YLYCDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 L167C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPC PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 82 
                 G-FGF21 H140C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 L142C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EACGCPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 83 
                 G-FGF21 D130C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 Y132C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LECGCNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 84 
                 G-FGF21 S137C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 H140C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQC EACGLPLHLPGNKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 85 
                 G-FGF21 S76C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQCP 
               
               
                   
                 S79C 
                 ECLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 86 
                 G-FGF21 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 G108C, L142C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD CALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGCPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 87 
                 G-FGF21 A72C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGCADQSP 
               
               
                   
                 S79C 
                 ECLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 88 
                 G-FGF21 H60C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EACLEIREDG TVGGAADQSP 
               
               
                   
                 S79C, H145C, 
                 ECLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 L165C 
                 LEDGYNVYQS EAHGLPLCLP GNKSPHRDPA PRGPARFCPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 89 
                 G-FGF21 A72C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGCADQSP 
               
               
                   
                 S79C, G108C, 
                 ECLLQLKALK PGVIQILGVK TSRFLCQRPD CALYGSLHFD PEACSFRELL 
               
               
                   
                 L142C 
                 LEDGYNVYQS EAHGCPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 90 
                 G-FGF21 R47C, 
                 GHPIPDSSPL LQFGGQVRQC YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 P174C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPACPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 91 
                 G-FGF21 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 M196C, R203C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSCV GPSQGCSPSY AS 
               
               
                   
               
               
                 92 
                 G-FGF21 D107C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 D155C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPC GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRCPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 93 
                 G-FGF21 D155C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDACQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 G148C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP CNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 94 
                 G-FGF21 Q550, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDACQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 G148C, N149- 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 R159 deleted and 
                 LEDGYNVYQS EAHGLPLHLP CGSGSGPARF LPLPGLPPAP PEPPGILAPQ 
               
               
                   
                 replaced by 
                 PPDVGSSDPL SMVGPSQGRS PSYAS 
               
               
                   
                 GSGS(SEQ ID 
                   
               
               
                   
                 NO: 161) 
                   
               
               
                   
               
               
                 95 
                 G-FGF21 Q55C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDACQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 G148C, N149- 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 R159 deleted and 
                 LEDGYNVYQS EAHGLPLHLP CGGSGGSGPA RFLPLPGLPP APPEPPGILA 
               
               
                   
                 replaced by 
                 PQPPDVGSSD PLSMVGPSQG RSPSYAS 
               
               
                   
                 GGSGGS (SEQ 
                   
               
               
                   
                 ID NO: 162) 
                   
               
               
                   
               
               
                 96 
                 G-FGF21 D155G, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAGCT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 Q560, L146C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHCP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 97 
                 G-FGF21 Q55G, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAGCT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 Q56C, L146C, 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 P147-R159 
                 LEDGYNVYQS EAHGLPLHCG SGSGPARFLP LPGLPPAPPE PPGILAPQPP 
               
               
                   
                 deleted and 
                 DVGSSDPLSM VGPSQGRSPS YAS 
               
               
                   
                 replaced by 
                   
               
               
                   
                 GSGS (SEQ ID 
                   
               
               
                   
                 NO: 161) 
                   
               
               
                   
               
               
                 98 
                 G-FGF21 Q55G, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAGCT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 Q56C, L146C, 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 P147-R159 
                 LEDGYNVYQS EAHGLPLHCG GSGGSGPARF LPLPGLPPAP PEPPGILAPQ 
               
               
                   
                 deleted and 
                 PPDVGSSDPL SMVGPSQGRS PSYAS 
               
               
                   
                 replaced by 
                   
               
               
                   
                 GGSGGS (SEQ 
                   
               
               
                   
                 ID NO: 162) 
                   
               
               
                   
               
               
                 99 
                 G-FGF21 Q55C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDACQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 P1470 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLC GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 100 
                 G-FGF21 Q55C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDACQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 N1490 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GCKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 101 
                 G-FGF21 E62C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLCIREDG TVGGAADQSP 
               
               
                   
                 E78C 
                 CSLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 102 
                 G-FGF21 E62C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLCIREDG TVGGAADQSP 
               
               
                   
                 S79C 
                 ECLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 103 
                 G-FGF21 R100C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 L114C 
                 ESLLQLKALK PGVIQILGVK TSCFLCQRPD GALYGSCHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 104 
                 G-FGF21 L102C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 G112C 
                 ESLLQLKALK PGVIQILGVK TSRFCCQRPD GALYCSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 105 
                 G-FGF21 A59C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT ECHLEIREDG TVGGACDQSP 
               
               
                   
                 A73C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 106 
                 G-FGF21 E62C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLCIREDG TVGGCADQSP 
               
               
                   
                 A72C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 107 
                 G-FGF21 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 Q104C, S137C 
                 ESLLQLKALK PGVIQILGVK TSRFLCCRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQC EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 108 
                 G-FGF21 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 Q104C, H140C 
                 ESLLQLKALK PGVIQILGVK TSRFLCCRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EACGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 109 
                 G-FGF21 A54C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDCQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 P161C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGCARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 110 
                 G-FGF21 Q56C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDACQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 L146C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPCHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 111 
                 G-FGF21 I63C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLECREDG TVGGAADQSP 
               
               
                   
                 T98C 
                 ESLLQLKALK PGVIQILGVK CSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 112 
                 G-FGF21 G67C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDC TVGGAADQSP 
               
               
                   
                 R100C 
                 ESLLQLKALK PGVIQILGVK TSCFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 113 
                 G-FGF21 V69C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TCGGAADQSP 
               
               
                   
                 L110C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GACYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 114 
                 G-FGF21 A120C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 A139C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PECCSFRELL 
               
               
                   
                   
                 LEDGYNVYQS ECHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 115 
                 G-FGF21 R124C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 Q136C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFCELL 
               
               
                   
                   
                 LEDGYNVYCS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 116 
                 G-FGF21 P143C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 A162C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLCLHLP GNKSPHRDPA PRGPCRFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 117 
                 G-FGF21 P77C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSC 
               
               
                   
                 K97C 
                 ESLLQLKALK PGVIQILGVC TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 118 
                 G-FGF21 E78C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 T98C 
                 CSLLQLKALK PGVIQILGVK CSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 119 
                 G-FGF21 L80C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 V96C 
                 ESCLQLKALK PGVIQILGCK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 120 
                 G-FGF21 H145C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 P161C 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLCLP GNKSPHRDPA PRGCARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 121 
                 G-FGF21 L81C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 G95C 
                 ESLCQLKALK PGVIQILCVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 122 
                 G-FGF21 Q82C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 L94C 
                 ESLLCLKALK PGVIQICGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 123 
                 G-FGF21 L83C, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 I91C 
                 ESLLQCKALK PGVCQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 124 
                 G-FGF21 D155L 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                   
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRLPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 125 
                 G-FGF21 D155K 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                   
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRKPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 126 
                 G-FGF21 D155Y 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                   
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRYPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 127 
                 G-FGF21 D155P 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                   
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRPPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 128 
                 G-FGF21 D155E 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                   
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHREPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 129 
                 G-FGF21 D155N 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                   
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRNPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 130 
                 G-FGF21 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 R154Q, D155L 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHQLPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 131 
                 G-FGF21 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 R154Q, D155K 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHQKPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 132 
                 G-FGF21 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 H153Q, R154Q, 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 D155N, A157V, 
                 LEDGYNVYQS EAHGLPLHLP GNKSPQQNPV PKGPARFLPL PGLPPAPPEP 
               
               
                   
                 R159K 
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 133 
                 G-FGF21 P152A, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 H153K, R154K, 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 D155E, P156A, 
                 LEDGYNVYQS EAHGLPLHLP GNKSAKKEAS PQGPARFLPL PGLPPAPPEP 
               
               
                   
                 A157S, R159Q 
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 134 
                 G-FGF21 H153Y, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 R154K, D155N, 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 P156K, A157G 
                 LEDGYNVYQS EAHGLPLHLP GNKSPYKNKG PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 135 
                 G-FGF21 P152- 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 H153 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSRDPAPR GPARFLPLPG LPPAPPEPPG 
               
               
                   
                   
                 ILAPQPPDVG SSDPLSMVGP SQGRSPSYAS 
               
               
                   
               
               
                 136 
                 G-FGF21 N149- 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 R163 deleted and 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 replaced by 
                 LEDGYNVYQS EAHGLPLHLP GGSGSFLPLP GLPPAPPEPP GILAPQPPDV 
               
               
                   
                 GSGS (SEQ ID 
                 GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
                 NO: 161) 
                   
               
               
                   
               
               
                 137 
                 G-FGF21 N149- 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 A162 deleted and 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 replaced by 
                 LEDGYNVYQS EAHGLPLHLP GGSHSGRFLP LPGLPPAPPE PPGILAPQPP 
               
               
                   
                 GSHSG (SEQ ID 
                 DVGSSDPLSM VGPSQGRSPS YAS 
               
               
                   
                 NO: 163) 
                   
               
               
                   
               
               
                 138 
                 G-FGF21 N149- 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 A162 deleted and 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 replaced by 
                 LEDGYNVYQS EAHGLPLHLP GGSHSGSRFL PLPGLPPAPP EPPGILAPQP 
               
               
                   
                 GSHSGS (SEQ 
                 PDVGSSDPLS MVGPSQGRSP SYAS 
               
               
                   
                 ID NO: 165) 
                   
               
               
                   
               
               
                 139 
                 G-FGF21 K150H 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                   
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNHSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 140 
                 G-FGF21 K150H, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 P152L 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNHSLHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 141 
                 G-FGF21 R163H 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                   
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPAHFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 142 
                 G-FGF21 P158H, 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 R159H 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA HHGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 143 
                 G-FGF21 N149- 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 D155 deleted and 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 replaced by 
                 LEDGYNVYQS EAHGLPLHLP GGSGSPAPRG PARFLPLPGL PPAPPEPPGI 
               
               
                   
                 GSGS (SEQ ID 
                 LAPQPPDVGS SDPLSMVGPS QGRSPSYAS 
               
               
                   
                 NO: 161) 
                   
               
               
                   
               
               
                 144 
                 G-FGF21 N149- 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 D155 deleted and 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 replaced by 
                 LEDGYNVYQS EAHGLPLHLP GGSHSGPAPR GPARFLPLPG LPPAPPEPP 
               
               
                   
                 GSHSG (SEQ ID 
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSY AS 
               
               
                   
                 NO: 163) 
                   
               
               
                   
               
               
                 145 
                 G-FGF21 N149- 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 D155 deleted and 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 replaced by 
                 LEDGYNVYQS EAHGLPLHLP GATTSPAPRG PARFLPLPGL PPAPPEPPGI 
               
               
                   
                 ATTS (SEQ ID 
                 LAPQPPDVGS SDPLSMVGPS QGRSPSYAS 
               
               
                   
                 NO: 164) 
                   
               
               
                   
               
               
                 146 
                 G-FGF21 R159- 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 R163 deleted and 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 replaced by GA 
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PGAFLPLPGL PPAPPEPPGI 
               
               
                   
                   
                 LAPQPPDVGS SDPLSMVGPS QGRSPSYAS 
               
               
                   
               
               
                 147 
                 G-FGF21 R159- 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 R163 deleted and 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 replaced by GY 
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PGYFLPLPGL PPAPPEPPGI 
               
               
                   
                   
                 LAPQPPDVGS SDPLSMVGPS QGRSPSYAS 
               
               
                   
               
               
                 148 
                 G-FGF21 R159- 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 R163 deleted and 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 replaced by HH 
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PHHFLPLPGL PPAPPEPPGI 
               
               
                   
                   
                 LAPQPPDVGS SDPLSMVGPS QGRSPSYAS 
               
               
                   
               
               
                 149 
                 G-FGF21 R159- 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 R163 deleted and 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 replaced by GE 
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPAPGEFLPLPGLPPAPPEPPGILA 
               
               
                   
                   
                 PQPPDVGSSD PLSMVGPSQG RSPSYAS 
               
               
                   
               
               
                 150 
                 G-FGF21 R159- 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 R163 deleted and 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 replaced by HE 
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PHEFLPLPGL PPAPPEPPGI 
               
               
                   
                   
                 LAPQPPDVGS SDPLSMVGPS QGRSPSYAS 
               
               
                   
               
               
                 151 
                 G-FGF21 A162Y 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                   
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPYRFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 152 
                 G-FGF21 R159H 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                   
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PHGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
               
               
                 153 
                 G-FGF21 5200M 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                   
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPMQGRSPSY AS 
               
               
                   
               
               
                 154 
                 G-FGF21 G2021 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                   
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQTRSPSY AS 
               
               
                   
               
               
                 155 
                 G-FGF21 R203E 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                   
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGESPSY AS 
               
               
                   
               
               
                 156 
                 G-FGF21 R203H 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                   
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQGHSPSY AS 
               
               
                   
               
               
                 157 
                 G-FGF21 G202- 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 R203 deleted 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                   
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                   
                 PGILAPQPPD VGSSDPLSMV GPSQSPSYAS 
               
               
                   
               
               
                 158 
                 FGF21 (S34- 
                 SSPLLQFGGQ VRQRYLYTDD ACQTEAHLEI REDGTVGGAA DQSPESLLQL 
               
               
                   
                 E176), Q55C, 
                 KALKPGVIQI LGVKTSRFLC QRPDGALYGS LHFDPEACSF RELLLEDGYN 
               
               
                   
                 N149C 
                 VYQSEAHGLP LHLPGCKSPH RDPAPRGPAR FLPLPGLPPA PPE 
               
               
                   
               
               
                 159 
                 G-FGF21 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 PLSMVGPSQGR 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 SPSYAS (SEQ 
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                 ID NO: 169) 
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY ASPLSMVGPS QGRSPSYAS 
               
               
                   
                 inserted after 
                   
               
               
                   
                 S209 
                   
               
               
                   
               
               
                 160 
                 G-FGF21 P199 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 deleted, 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 PLSMVGSQGRS 
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPAPPEP 
               
               
                   
                 PSYAS(SEQ ID 
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY ASPLSMVGSQ GRSPSYAS 
               
               
                   
                 NO: 170) inserted 
                   
               
               
                   
                 after S209 
                   
               
               
                   
               
               
                 161 
                 Linker 
                 GSGS 
               
               
                   
               
               
                 162 
                 Linker 
                 GGSGGS 
               
               
                   
               
               
                 163 
                 Linker 
                 GSHSG 
               
               
                   
               
               
                 164 
                 Linker 
                 ATTS 
               
               
                   
               
               
                 165 
                 Linker 
                 GSHSGS 
               
               
                   
               
               
                 166 
                 Linker 
                 GHRSHLQTVF 
               
               
                   
               
               
                 167 
                 Linker 
                 GLNSMV 
               
               
                   
               
               
                 168 
                 Linker 
                 GGGGS 
               
               
                   
               
               
                 169 
                 C-terminal 
                 PLSMVGPSQG RSPSYAS 
               
               
                   
                 extension 
                   
               
               
                   
               
               
                 170 
                 C-terminal 
                 PLSMVGSQGR SPSYAS 
               
               
                   
                 extension 
                   
               
               
                   
               
               
                 171 
                 FGF21 variant, 
                 MDSDETGFEH SGLWVSVLAG LLLGACQAHP IPDSSPLLQF GGQVRQRYLY 
               
               
                   
                 consensus 
                 TDDAX 55 QTEAH LEIREDGTVG GAADQSPESL LQLKALKPGV IQILGVKTSR 
               
               
                   
                 sequence I 
                 FLCQRPDGAL YGSLHFDPEA CSFRELLLED GYNVYQSEAH 
               
               
                   
                   
                 GLPLHLX 147 X 148 X 149 X 150  X 151 X 152 X 153 X 154 X 155 X 156 X 157 X 158 X 159 X 160 X 161   
               
               
                   
                   
                 X 162 X 163FLPLPGL PPAPPEPPGI LAP X 184 X 185 X 186 X 187 X 188 X 189 X 190   
               
               
                   
                   
                 X 191 X 192 X 193 X 194 X 195 X 196 X 197 X 198 X 199 X 200  X 201 X 202  X 203 X 204 X 205 X 206 X 207 AS 
               
               
                   
               
               
                 172 
                 FGF21 S141, 
                 MDSDETGFEH SGLWVSVLAG LLLGACQAHP IPDSSPLLQF GGQVRQRYLY 
               
               
                   
                 natural variant 
                 TDDAQQTEAH LEIREDGTVG GAADQSPESL LQLKALKPGV IQILGVKTSR 
               
               
                   
                   
                 FLCQRPDGAL YGSLHFDPEA CSFRELLLED GYNVYQSEAH SLPLHLPGNK 
               
               
                   
                   
                 SPHRDPAPRG PARFLPLPGL PPAPPEPPGI LAPQPPDVGS SDPLSMVGPS 
               
               
                   
                   
                 QGRSPSYAS 
               
               
                   
               
               
                 173 
                 FGF21 L174, 
                 MDSDETGFEH SGLWVSVLAG LLLGACQAHP IPDSSPLLQF GGQVRQRYLY 
               
               
                   
                 natural variant 
                 TDDAQQTEAH LEIREDGTVG GAADQSPESL LQLKALKPGV IQILGVKTSR 
               
               
                   
                   
                 FLCQRPDGAL YGSLHFDPEA CSFRELLLED GYNVYQSEAH GLPLHLPGNK 
               
               
                   
                   
                 SPHRDPAPRG PARFLPLPGL PPALPEPPGI LAPQPPDVGS SDPLSMVGPS 
               
               
                   
                   
                 QGRSPSYAS 
               
               
                   
               
               
                 174 
                 mature human 
                 GHPIPDSSPL LQFGGQVRQR YLYTDDAQQT EAHLEIREDG TVGGAADQSP 
               
               
                   
                 wild-type FGF21 
                 ESLLQLKALK PGVIQILGVK TSRFLCQRPD GALYGSLHFD PEACSFRELL 
               
               
                   
                 variant L174 with  
                 LEDGYNVYQS EAHGLPLHLP GNKSPHRDPA PRGPARFLPL PGLPPALPEP 
               
               
                   
                 additional N- 
                 PGILAPQPPD VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
                 terminal G (G- 
                   
               
               
                   
                 FGF21) 
                   
               
               
                   
               
               
                 175 
                 FGF21 variant, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAX 55 QTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 consensus 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 sequence II 
                 EDGYNVYQSE AHGL2LHLX 147 X 148  X 149 KSPHRX 155 PAP RGX 161 X 162 RFLPLP 
               
               
                   
                   
                 GLPPAPPEPP GILAPQPPDV GSSDPLX 195 X 196 X 197 X 198   
               
               
                   
                   
                 X 199 X 200 X 201 X 202 X 203 SPSYA S 
               
               
                   
               
               
                 176 
                 FGF21 variant, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAX 55 QTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 consensus 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 sequence III 
                 EDGYNVYQSE AHGLPLHLX 147 X 148  X 149 KSPHRX 155 PAP RGPARFLPLP 
               
               
                   
                   
                 GLPPAPPEPP GILAPQPPDV GSSDPLSMVX 198  X l99 SQGRSPSYA S 
               
               
                   
               
               
                 177 
                 FGF21 S195C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 S200C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLCMVG PCQGRSPSYA S 
               
               
                   
               
               
                 178 
                 FGF21 P199S, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 S200P 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG SPQGRSPSYA S 
               
               
                   
               
               
                 179 
                 FGF21 P199Q, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 Q201P 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG QSPGRSPSYA S 
               
               
                   
               
               
                 180 
                 FGF21 P199 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG SQGRSPSYAS 
               
               
                   
               
               
                 181 
                 FGF21 M196P, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 V197G, G198V, 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 P199M 
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSPGV MSQGRSPSYA S 
               
               
                   
               
               
                 182 
                 FGF21 M196V, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 V197M 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSVMG PSQGRSPSYA S 
               
               
                   
               
               
                 183 
                 FGF21 V197E 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                   
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMEG PSQGRSPSYA S 
               
               
                   
               
               
                 184 
                 FGF21 GGGGS 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 (SEQ ID NO: 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 168) inserted 
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                 between G198 
                 GILAPQPPDV GSSDPLSMVG GGGGSPSQGR SPSYAS 
               
               
                   
                 and P199 
                   
               
               
                   
               
               
                 185 
                 FGF21 G198E 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                   
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVE PSQGRSPSYA S 
               
               
                   
               
               
                 186 
                 FGF21 G198C 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                   
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVD PSQGRSPSYA S 
               
               
                   
               
               
                 187 
                 FGF21 G198R 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                   
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVR PSQGRSPSYA S 
               
               
                   
               
               
                 188 
                 FGF21 G198K 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                   
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVK PSQGRSPSYA S 
               
               
                   
               
               
                 189 
                 FGF21 G198Y 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                   
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVY PSQGRSPSYA S 
               
               
                   
               
               
                 190 
                 FGF21 P199G 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                   
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG GSQGRSPSYA S 
               
               
                   
               
               
                 191 
                 FGF21 P199R 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                   
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG RSQGRSPSYA S 
               
               
                   
               
               
                 192 
                 FGF21 P1991 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                   
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG TSQGRSPSYA S 
               
               
                   
               
               
                 193 
                 FGF21 G198P, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 P199G 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVP GSQGRSPSYA S 
               
               
                   
               
               
                 194 
                 FGF21 S200Q, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 Q201S 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PQSGRSPSYA S 
               
               
                   
               
               
                 195 
                 FGF21 V197C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 G198V, P199C, 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 S200H 
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMDV DHQGRSPSYA S 
               
               
                   
               
               
                 196 
                 FGF21 V197- 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 R203 deleted and 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 replaced by 
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                 GHRSHLQTVF 
                 GILAPQPPDV GSSDPLSMGH RSHLQTVFSP SYAS 
               
               
                   
                 (SEQ ID NO: 166) 
                   
               
               
                   
               
               
                 197 
                 FGF21 V197- 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 R203 deleted and 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 replaced by 
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                 GLNSMV (SEQ 
                 GILAPQPPDV GSSDPLSMGL NSMVSPSYAS 
               
               
                   
                 ID NO: 167) 
                   
               
               
                   
               
               
                 198 
                 FGF21 G198 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVP SQGRSPSYAS 
               
               
                   
               
               
                 199 
                 FGF21 G198- 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 P199 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVS QGRSPSYAS 
               
               
                   
               
               
                 200 
                 FGF21 G198 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted, P199F 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVF SQGRSPSYAS 
               
               
                   
               
               
                 201 
                 FGF21 G198 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted, P199L 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVL SQGRSPSYAS 
               
               
                   
               
               
                 202 
                 FGF21 G198 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted, P199L, 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 S200N 
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVL NQGRSPSYAS 
               
               
                   
               
               
                 203 
                 FGF21 V197 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMGP SQGRSPSYAS 
               
               
                   
               
               
                 204 
                 FGF21 L146C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 A162C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHCPG NKSPHRDPAP RGPCRFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 205 
                 FGF21 A59C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE CHLEIREDGT VGCAADQSPE 
               
               
                   
                 G71C, P199 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 deleted 
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG SQGRSPSYAS 
               
               
                   
               
               
                 206 
                 FGF21 A59C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE CHLEIREDGT VGCAADQSPE 
               
               
                   
                 G71C, G198Y 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVY PSQGRSPSYA S 
               
               
                   
               
               
                 207 
                 FGF21 S76C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQCPE 
               
               
                   
                 S79C, P199 
                 CLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 deleted 
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG SQGRSPSYAS 
               
               
                   
               
               
                 208 
                 FGF21 S76C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQCPE 
               
               
                   
                 S79C, G198Y 
                 CLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVY PSQGRSPSYA S 
               
               
                   
               
               
                 209 
                 FGF21 G108C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 L142C, P199 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDC ALYGSLHFDP EACSFRELLL 
               
               
                   
                 deleted 
                 EDGYNVYQSE AHGCPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG SQGRSPSYAS 
               
               
                   
               
               
                 210 
                 FGF21 G108C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 L142C, G198Y 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDC ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGCPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVY PSQGRSPSYA S 
               
               
                   
               
               
                 211 
                 FGF21 D107C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 D155C, P199 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPCG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 deleted 
                 EDGYNVYQSE AHGLPLHLPG NKSPHRCPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG SQGRSPSYAS 
               
               
                   
               
               
                 212 
                 FGF21 D107C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 D155C, G198Y 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPCG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRCPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVY PSQGRSPSYA S 
               
               
                   
               
               
                 213 
                 FGF21 Q55C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDACQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 P147C, P199 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 deleted 
                 EDGYNVYQSE AHGLPLHLCG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG SQGRSPSYAS 
               
               
                   
               
               
                 214 
                 FGF21 Q55C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDACQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 P147C, G198Y 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLCG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVY PSQGRSPSYA S 
               
               
                   
               
               
                 215 
                 FGF21 Q55C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDACQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 N1490, P199 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 deleted 
                 EDGYNVYQSE AHGLPLHLPG CKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG SQGRSPSYAS 
               
               
                   
               
               
                 216 
                 FGF21 Q55C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDACQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 N149C, G198Y 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG CKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVY PSQGRSPSYA S 
               
               
                   
               
               
                 217 
                 FGF21 M196 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSVGP SQGRSPSYAS 
               
               
                   
               
               
                 218 
                 FGF21 S195 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLMVGP SQGRSPSYAS 
               
               
                   
               
               
                 219 
                 FGF21 L194 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPSMVGP SQGRSPSYAS 
               
               
                   
               
               
                 220 
                 FGF21 P193 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDLSMVGP SQGRSPSYAS 
               
               
                   
               
               
                 221 
                 FGF21 D192 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSPLSMVGP SQGRSPSYAS 
               
               
                   
               
               
                 222 
                 FGF21 S191 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSDPLSMVGP SQGRSPSYAS 
               
               
                   
               
               
                 223 
                 FGF21 S190- 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 S191 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GDPLSMVGPS QGRSPSYAS 
               
               
                   
               
               
                 224 
                 FGF21 G189 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV SSDPLSMVGP SQGRSPSYAS 
               
               
                   
               
               
                 225 
                 FGF21 V188 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDG SSDPLSMVGP SQGRSPSYAS 
               
               
                   
               
               
                 226 
                 FGF21 D187 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPVG SSDPLSMVGP SQGRSPSYAS 
               
               
                   
               
               
                 227 
                 FGF21 P186 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPDVG SSDPLSMVGP SQGRSPSYAS 
               
               
                   
               
               
                 228 
                 FGF21 P185- 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 P186 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQDVGS SDPLSMVGPS QGRSPSYAS 
               
               
                   
               
               
                 229 
                 FGF21 Q184 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPPPDVG SSDPLSMVGP SQGRSPSYAS 
               
               
                   
               
               
                 230 
                 FGF21 S200 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PQGRSPSYAS 
               
               
                   
               
               
                 231 
                 FGF21 Q201 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSGRSPSYAS 
               
               
                   
               
               
                 232 
                 FGF21 G202 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQRSPSYAS 
               
               
                   
               
               
                 233 
                 FGF21 R203 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGSPSYAS 
               
               
                   
               
               
                 234 
                 FGF21 S204 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRPSYAS 
               
               
                   
               
               
                 235 
                 FGF21 P205 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSSYAS 
               
               
                   
               
               
                 236 
                 FGF21 S206 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPYAS 
               
               
                   
               
               
                 237 
                 FGF21 Y207 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSAS 
               
               
                   
               
               
                 238 
                 FGF21 V197 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted, S201 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 deleted 
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMGP QGRSPSYAS 
               
               
                   
               
               
                 239 
                 FGF21 M196 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted, S201 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 deleted 
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSVGP QGRSPSYAS 
               
               
                   
               
               
                 240 
                 FGF21 M196- 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 V197 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSGPS QGRSPSYAS 
               
               
                   
               
               
                 241 
                 FGF21 V197- 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 G198 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMPS QGRSPSYAS 
               
               
                   
               
               
                 242 
                 FGF21 S195- 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 M196 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLVGPS QGRSPSYAS 
               
               
                   
               
               
                 243 
                 FGF21 G198 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted, S200 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 deleted 
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVP QGRSPSYAS 
               
               
                   
               
               
                 244 
                 FGF21 H60C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE ACLEIREDGT VGGAADQSPE 
               
               
                   
                 S79C 
                 CLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 245 
                 FGF21 H145C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 L165C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLCLPG NKSPHRDPAP RGPARFCPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 246 
                 FGF21 A59C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE CHLEIREDGT VGCAADQSPE 
               
               
                   
                 G71C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 247 
                 FGF21 H60C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE ACLEIREDGT VGGCADQSPE 
               
               
                   
                 A72C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 248 
                 FGF21 I63C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLECREDGT VGGAADQSPE 
               
               
                   
                 G112C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYCSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 249 
                 FGF21 L49C, 
                 HPIPDSSPLL QFGGQVRQRY CYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 L170C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GCPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 250 
                 FGF21 151C, 
                 HPIPDSSPLL QFGGQVRQRY LYCDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 L167C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPCP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 251 
                 FGF21 H140C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 L142C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE ACGCPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                 252 
                 FGF21 D130C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 Y132C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 ECGCNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                 253 
                 FGF21 S137C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 H140C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQCE ACGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 254 
                 FGF21 S76C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQCPE 
               
               
                   
                 S79C 
                 CLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                 255 
                 FGF21 G108C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 L142C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDC ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGCPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 256 
                 FGF21 A72C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGCADQSPE 
               
               
                   
                 S79C 
                 CLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 257 
                 FGF21 H60C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE ACLEIREDGT VGGAADQSPE 
               
               
                   
                 S79C, H145C, 
                 CLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 L165C 
                 EDGYNVYQSE AHGLPLCLPG NKSPHRDPAP RGPARFCPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 258 
                 FGF21 A72C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGCADQSPE 
               
               
                   
                 S79C, G108C, 
                 CLLQLKALKP GVIQILGVKT SRFLCQRPDC ALYGSLHFDP EACSFRELLL 
               
               
                   
                 L142C 
                 EDGYNVYQSE AHGCPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 259 
                 FGF21 R47C, 
                 HPIPDSSPLL QFGGQVRQCY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 P174C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPACPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 260 
                 FGF21 M196C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 R203C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSCVG PSQGCSPSYA S 
               
               
                   
               
               
                 261 
                 FGF21 D107C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 D155C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPCG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRCPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 262 
                 FGF21 Q55C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDACQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 G148C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPC NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 263 
                 FGF21 Q55C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDACQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 G148C, N149- 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 R159 deleted and 
                 EDGYNVYQSE AHGLPLHLPC GSGSGPARFL PLPGLPPAPP EPPGILAPQP 
               
               
                   
                 replaced by 
                 PDVGSSDPLS MVGPSQGRSP SYAS 
               
               
                   
                 GSGS(SEQ ID 
                   
               
               
                   
                 NO: 161) 
                   
               
               
                   
               
               
                 264 
                 FGF21 Q55C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDACQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 G148C, N149- 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 R159 deleted and 
                 EDGYNVYQSE AHGLPLHLPC GGSGGSGPAR FLPLPGLPPA PPEPPGILAP 
               
               
                   
                 replaced by 
                 QPPDVGSSDP LSMVGPSQGR SPSYAS 
               
               
                   
                 GGSGGS (SEQ 
                   
               
               
                   
                 ID NO: 162) 
                   
               
               
                   
               
               
                 265 
                 FGF21 Q55G, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAGCTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 Q56C, L146C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHCPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 266 
                 FGF21 Q55G, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAGCTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 Q56C, L146C, 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 P147-R159 
                 EDGYNVYQSE AHGLPLHCGS GSGPARFLPL PGLPPAPPEP PGILAPQPPD 
               
               
                   
                 deleted and 
                 VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
                 replaced by 
                   
               
               
                   
                 GSGS (SEQ ID 
                   
               
               
                   
                 NO: 161) 
                   
               
               
                   
               
               
                 267 
                 FGF21 Q55G, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAGCTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 Q56C, L146C, 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 P147-R159 
                 EDGYNVYQSE AHGLPLHCGG SGGSGPARFL PLPGLPPAPP EPPGILAPQP 
               
               
                   
                 deleted and 
                 PDVGSSDPLS MVGPSQGRSP SYAS 
               
               
                   
                 replaced by 
                   
               
               
                   
                 GGSGGS (SEQ 
                   
               
               
                   
                 ID NO: 162) 
                   
               
               
                   
               
               
                 268 
                 FGF21 Q55C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDACQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 P147C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLCG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 269 
                 FGF21 Q55C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDACQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 N149C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG CKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 270 
                 FGF21 E62C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLCIREDGT VGGAADQSPC 
               
               
                   
                 E78C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 271 
                 FGF21 E62C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLCIREDGT VGGAADQSPE 
               
               
                   
                 S79C 
                 CLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 272 
                 FGF21 R100C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 L114C 
                 SLLQLKALKP GVIQILGVKT SCFLCQRPDG ALYGSCHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 273 
                 FGF21 L102C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 G112C 
                 SLLQLKALKP GVIQILGVKT SRFCCQRPDG ALYCSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 274 
                 FGF21 A59C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE CHLEIREDGT VGGACDQSPE 
               
               
                   
                 A73C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 275 
                 FGF21 E62C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLCIREDGT VGGCADQSPE 
               
               
                   
                 A72C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 276 
                 FGF21 Q104C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 S137C 
                 SLLQLKALKP GVIQILGVKT SRFLCCRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQCE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 277 
                 FGF21 Q104C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 H140C 
                 SLLQLKALKP GVIQILGVKT SRFLCCRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE ACGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 278 
                 FGF21 A54C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDCQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 P161C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGCARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 279 
                 FGF21 Q56C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDACQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 L146C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPCHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 280 
                 FGF21 I63C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLECREDGT VGGAADQSPE 
               
               
                   
                 T98C 
                 SLLQLKALKP GVIQILGVKC SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 281 
                 FGF21 G67C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDCT VGGAADQSPE 
               
               
                   
                 R100C 
                 SLLQLKALKP GVIQILGVKT SCFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 282 
                 FGF21 V69C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT CGGAADQSPE 
               
               
                   
                 L110C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ACYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 283 
                 FGF21 A120C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 A139C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP ECCSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE CHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 284 
                 FGF21 R124C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 Q136C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFCELLL 
               
               
                   
                   
                 EDGYNVYCSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 285 
                 FGF21 P143C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 A162C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLCLHLPG NKSPHRDPAP RGPCRFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 286 
                 FGF21 P77C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSCE 
               
               
                   
                 K97C 
                 SLLQLKALKP GVIQILGVCT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 287 
                 FGF21 E78C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPC 
               
               
                   
                 T98C 
                 SLLQLKALKP GVIQILGVKC SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 288 
                 FGF21 L80C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 V96C 
                 SCLQLKALKP GVIQILGCKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 289 
                 FGF21 H145C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 P161C 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLCLPG NKSPHRDPAP RGCARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 290 
                 FGF21 L81C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 G95C 
                 SLCQLKALKP GVIQILCVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 291 
                 FGF21 Q82C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 L94C 
                 SLLCLKALKP GVIQICGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 292 
                 FGF21 L83C, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 I91C 
                 SLLQCKALKP GVCQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 293 
                 FGF21 D155L 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                   
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRLPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 294 
                 FGF21 D155K 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                   
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRKPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 295 
                 FGF21 D155Y 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                   
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRYPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 296 
                 FGF21 D155P 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                   
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRPPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 297 
                 FGF21 D155E 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                   
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHREPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 298 
                 FGF21 D155N 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                   
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRNPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 299 
                 FGF21 R154Q, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 D155L 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHQLPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 300 
                 FGF21 R154Q, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 D155K 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHQKPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 301 
                 FGF21 H153Q, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 R154Q, D155N, 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 A157V, R159K 
                 EDGYNVYQSE AHGLPLHLPG NKSPQQNPVP KGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 302 
                 FGF21 P152A, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 H153K, R154K, 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 D155E, P156A, 
                 EDGYNVYQSE AHGLPLHLPG NKSAKKEASP QGPARFLPLP GLPPAPPEPP 
               
               
                   
                 A157S, R159Q 
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
                   
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
               
               
                 303 
                 FGF21 H153Y, 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 R154K, D155N, 
                 EDGYNVYQSE AHGLPLHLPG NKSPYKNKGP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                 P156K, A157G 
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 304 
                 FGF21 P152- 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 H153 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSRDPAPRG PARFLPLPGL PPAPPEPPGI 
               
               
                   
                   
                 LAPQPPDVGS SDPLSMVGPS QGRSPSYAS 
               
               
                   
               
               
                 305 
                 FGF21 N149- 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 R163 deleted and 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 replaced by 
                 EDGYNVYQSE AHGLPLHLPG GSGSFLPLPG LPPAPPEPPG ILAPQPPDVG 
               
               
                   
                 GSGS (SEQ ID 
                 SSDPLSMVGP SQGRSPSYAS 
               
               
                   
                 NO: 161) 
                   
               
               
                   
               
               
                 306 
                 FGF21 N149- 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 A162 deleted and 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 replaced by 
                 EDGYNVYQSE AHGLPLHLPG GSHSGRFLPL PGLPPAPPEP PGILAPQPPD 
               
               
                   
                 GSHSG (SEQ ID 
                 VGSSDPLSMV GPSQGRSPSY AS 
               
               
                   
                 NO: 163) 
                   
               
               
                   
               
               
                 307 
                 FGF21 N149- 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 A162 deleted and 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 replaced by 
                 EDGYNVYQSE AHGLPLHLPG GSHSGSRFLP LPGLPPAPPE PPGILAPQPP 
               
               
                   
                 GSHSGS (SEQ 
                 DVGSSDPLSM VGPSQGRSPS YAS 
               
               
                   
                 ID NO: 165) 
                   
               
               
                   
               
               
                 308 
                 FGF21 K150H 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                   
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NHSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 309 
                 FGF21 K150H, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 P152L 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NHSLHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 310 
                 FGF21 R163H 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                   
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPAHFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 311 
                 FGF21 P158H, 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 R159H 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAH HGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 312 
                 FGF21 N149- 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 D155 deleted and 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 replaced by 
                 EDGYNVYQSE AHGLPLHLPG GSGSPAPRGP ARFLPLPGLP PAPPEPPGIL 
               
               
                   
                 GSGS (SEQ ID 
                 APQPPDVGSS DPLSMVGPSQ GRSPSYAS 
               
               
                   
                 NO: 161) 
                   
               
               
                   
               
               
                 313 
                 FGF21 N149- 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 D155 deleted and 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 replaced by 
                 EDGYNVYQSE AHGLPLHLPG GSHSGPAPRG PARFLPLPGL PPAPPEPPGI 
               
               
                   
                 GSHSG (SEQ ID 
                 LAPQPPDVGS SDPLSMVGPS QGRSPSYAS 
               
               
                   
                 NO: 163) 
                   
               
               
                   
               
               
                 314 
                 FGF21 N149- 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 D155 deleted and 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 replaced by 
                 EDGYNVYQSE AHGLPLHLPG ATTSPAPRGP ARFLPLPGLP PAPPEPPGIL 
               
               
                   
                 ATTS (SEQ ID 
                 APQPPDVGSS DPLSMVGPSQ GRSPSYAS 
               
               
                   
                 NO: 164) 
                   
               
               
                   
               
               
                 315 
                 FGF21 R159- 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 R163 deleted and 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 replaced by GA 
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP GAFLPLPGLP PAPPEPPGIL 
               
               
                   
                   
                 APQPPDVGSS DPLSMVGPSQ GRSPSYAS 
               
               
                   
               
               
                 316 
                 FGF21 R159- 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 R163 deleted and 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 replaced by GY 
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP GYFLPLPGLP PAPPEPPGIL 
               
               
                   
                   
                 APQPPDVGSS DPLSMVGPSQ GRSPSYAS 
               
               
                   
               
               
                 317 
                 FGF21 R159- 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 R163 deleted and 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 replaced by HH 
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP HHFLPLPGLP PAPPEPPGIL 
               
               
                   
                   
                 APQPPDVGSS DPLSMVGPSQ GRSPSYAS 
               
               
                   
               
               
                 318 
                 FGF21 R159- 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 R163 deleted and 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 replaced by GE 
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP GEFLPLPGLP PAPPEPPGIL 
               
               
                   
                   
                 APQPPDVGSS DPLSMVGPSQ GRSPSYAS 
               
               
                   
               
               
                 319 
                 FGF21 R159- 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 R163 deleted and 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 replaced by HE 
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP HEFLPLPGLP PAPPEPPGIL 
               
               
                   
                   
                 APQPPDVGSS DPLSMVGPSQ GRSPSYAS 
               
               
                   
               
               
                 320 
                 FGF21 A162Y 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                   
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPYRFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 321 
                 FGF21 R159H 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                   
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP HGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA S 
               
               
                   
               
               
                 322 
                 FGF21 S200M 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                   
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PMQGRSPSYA S 
               
               
                   
               
               
                 323 
                 FGF21 G2021 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                   
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQTRSPSYA S 
               
               
                   
               
               
                 324 
                 FGF21 R203E 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                   
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGESPSYA S 
               
               
                   
               
               
                 325 
                 FGF21 R203H 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                   
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQGHSPSYA S 
               
               
                   
               
               
                 326 
                 FGF21 G202- 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 R203 deleted 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                   
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                   
                 GILAPQPPDV GSSDPLSMVG PSQSPSYAS 
               
               
                   
               
               
                 327 
                 FGF21 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 PLSMVGPSQGR 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 SPSYAS (SEQ 
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                 ID NO: 169) 
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA SPLSMVGPSQ GRSPSYAS 
               
               
                   
                 inserted after 
                   
               
               
                   
                 S209 
                   
               
               
                   
               
               
                 328 
                 FGF21 P199 
                 HPIPDSSPLL QFGGQVRQRY LYTDDAQQTE AHLEIREDGT VGGAADQSPE 
               
               
                   
                 deleted, 
                 SLLQLKALKP GVIQILGVKT SRFLCQRPDG ALYGSLHFDP EACSFRELLL 
               
               
                   
                 PLSMVGSQGRS 
                 EDGYNVYQSE AHGLPLHLPG NKSPHRDPAP RGPARFLPLP GLPPAPPEPP 
               
               
                   
                 PSYAS(SEQ ID 
                 GILAPQPPDV GSSDPLSMVG PSQGRSPSYA SPLSMVGSQG RSPSYAS 
               
               
                   
                 NO:170) inseded 
                   
               
               
                   
                 after S209