Patent SG-11201903550T-A (SG)
Title: Protein screening and detection method
Inventor: SEEGER MARKUS
IPC: C07K16/12
Filed: 2017-10-30
Abstract: FigA Nanobody Library Nested Library (pNLx) Deepsequencing Vector (pNLs) Nested Library with Deepsequencing Adapters Flycode Library (pNLx) Vector exchange without PCR Restriction digest Isolated Flycodes E do 4 0 I Expression Oc\" Purification (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (43) International Publication Date 03 May 2018 (03.05.2018) WIP0 I PCT 1E1010 VIII °nolo III o Oil ioo Imo (10) International Publication Number WO 2018/078167 Al (51) International Patent Classification: C12N 15/10 (2006.01) G01N 33/68 (2006.01) C12N 15/62 (2006.01) C07K 16/12 (2006.01) (21) International Application Number: (22) International Filing Date: PCT/EP2017/077816 30 October 2017 (30.10.2017) (25) Filing Language: English (26) Publication Language: English (30) Priority Data: 16196571.0 31 October 2016 (31.10.2016) EP (71) Applicant: UNIVERSITAT ZURICH [CH/CH]; Prorek- torat VNW, Ramistr. 71, 8006 Ziirich (CH). (72) Inventors: SEEGER, Markus; Grundsteinweg 5, 8800 Thalwil (CH). EGLOFF, Pascal; Glamischstr. 5, 8800 Thalwil (CH). ZIMMERMANN, Iwan; Giblenstr. 27, 8049 Ziirich (CH). (74) Agent: HEINZ, Benjamin; Schulz Junghans Patentan- walte PartGmbB, GroBbeerenstrasse 71, 10963 Berlin (DE). (81) Designated States (unless otherwise indicated, for every kind of national protection available): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (84) Designated States (unless otherwise indicated, for every kind of regional protection available): ARIPO (BW, GH, (54) Title: PROTEIN SCREENING AND DETECTION METHOD Deepsequencing Massspectrometry NestLink expression vector backbone (grey) Adapters for Deepsequencing (black) B Nested Library (pNLx) Nanobodies Containing Unique Flycodes Selection Pressure Elution Volume (57) : The invention relates to a method for identifying and quantifying a polypeptide from a library of polypeptides. The method comprises the steps of: 1 - providing a polypeptide library and a detection tag library, 2 - generating a nested library comprising the polypeptides and the detection tags, 3 - sequencing the nested library, 4 - selecting a member of the nested library in one or several selection steps that are independent of a physical genotype-phenotype linkage, 5 - isolating the detection tag from the selected polypep- tide, 6 - identifying and quantifying the detection tag by mass spectrometry, 7 - obtaining the sequence of the selected polypeptide. The invention also relates to a collection of polypeptides, a collection of detection tags, and a collection of plasmid vectors. [Continued on next page] WO 2018/078167 Al MIDEDIMOMOIDEIREIDIONOMHOHINHIMEN GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG). Declarations under Rule 4.17: as to the identity of the inventor (Rule 4.17(0) as to applicant's entitlement to apply for and be granted a patent (Rule 4.170) Published: with international search report (Art. 21(3)) before the expiration of the time limit for amending the claims and to be republished in the event of receipt of amendments (Rule 48.2(h)) with sequence listing part of description (Rule 5.2(a))