Patent ID: 9383367
Filing Date: 2016-07-05
CPC Classification: C07K,C12Q,G01N

Claim Text:
1. A method of detecting a hydrolysis-created conjugation site-specific Artificially Cleaved Epitope (ACE) structure in a sample, wherein the ACE structure is hidden in its intact or natural form in a protein and is poorly accessible to antibodies, comprising steps of: (i) designing an ACE structure of formula Ln---L2-L1-(S1-S2---Sm)-L1′-L2′---Lm′ wherein the ACE structure consists of a first oligomer having the sequence Ln---L2-L1-L1′-L2′---Lm′, a second oligomer having the sequence S1-S2---Sm; wherein the second oligomer is covalently conjugated to the L1 residue via the S1 residue; wherein one or any combination of Ln, Lm′ and Sm are terminal residues that are artificially created via chemical bond-specific cleavage by at least one hydrolytic enzyme or hydrolytic agent; (ii) synthesizing the ACE structure; (iii) making an antibody against the ACE structure, wherein the antibody specifically recognizes both the first oligomer and the second oligomer; (iv) creating the ACE structure in the sample by treating the sample with the hydrolytic enzyme or hydrolytic agent, thereby exposing the formerly hidden ACE structure to specific interaction with the antibody; and (v) detecting the ACE structure created in step (iv) with the antibody, wherein the antibody specifically binds to both the first oligomer and the second oligomer.