Patent SG-11201809959P-A (SG)
Title: Methods of decreasing trisulfide bonds during recombinant production of polypeptides
Inventor: GAWLITZEK MARTIN
IPC: C12P21/08
Filed: 2017-05-09
Abstract: INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property 111111111111111111111111111111111111111111111111111111111111111111111111111111111111 Organization International Bureau (10) International Publication Number (43) International Publication Date .....0\"\" WO 2017/196902 A3 16 November 2017 (16.11.2017) WIPO I PCT (51) International Patent Classification: (72) Inventors: GAWLITZEK, Martin; c/o Genentech, Inc., 1 C12P 21/08 (2006.01) DNA Way, South San Francisco, CA 94080 (US). MARK- ERT, Sven; Penzberger Str. 16, 82404 Sindelsdorf (DE). (21) International Application Number: POPP, Oliver; Floesserstr. 15, 82377 Penzberg (DE). PCT/US2017/031832 SHIRATORI, Masaru, Ken; c/o Genentech, Inc., 1 Dna (22) International Filing Date: Way, South San Francisco, CA 94080 (US). TROBES, 09 May 2017 (09.05.2017) Thomas; Demleitnerstr. 23, 81371 Muenchen (DE). WUU, Jessica; c/o Genentech, Inc., 1 DNA Way, South San Fran- (25) Filing Language: English cisco, CA 94080 (US). (26) Publication Language: English (74) Agent: ONUFRYK, Christina et al.; Morrison & Foerster (30) Priority Data: LLP, 755 Page Mill Road, Palo Alto, CA 94304-1018 (US). 62/334,433 10 May 2016 (10.05.2016) US (81) Designated States (unless otherwise indicated, for every (71) Applicant (for all designated States except AL, AT, BA, kind of national protection available): AE, AG, AL, AM, BE, BG, CH, CN, CY, CZ, DE, DK, EE, ES, FI, FR, GB, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, GR, HR, HU, IE, IN, IS, IT, LT, LU, LV, MC, MK, MT, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR): GENEN- DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, TECH, INC. [US/US]; 1 DNA Way, South San Francisco, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KH, KN, KP, KR, CA 94080-4990 (US). 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, (71) Applicant (for AL, AT, BE, BG, CH, CN, CY, CZ, DE, DK, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, EE, ES, FI, FR, GB, GR, HR, HU, IE, IN, IS, IT, LT, LU, LV, = SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, = MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. only): F. HOFFMANN-LA ROCHE AG [CH/CH]; Gren- = zacherstrasse 124, 4070 Basel (CH). = = Title: METHODS OF DECREASING TRISULFIDE BONDS DURING RECOMBINANT PRODUCTION OF POLYPEPTIDES (54) FIG. 1 = = = 50 = = _ 40 _ = < —F—Media 1, cysteine + Fe E —II—Media 1, cystine + Fe = E 30 ' —O-Media 2, cysteine + Fe = —0—Media 2, cystine, + Fe = Z 7.—Media 1, cysteine 20 • —0—Media 1, cystine —c—Media 2, cysteine en 10 —#— Media 2, cystine ei © 0 irr • cT 0 12 24 36 48 60 72 t:T 1-1 Time (hours) Ir - - 1-1 N (57) : Provided herein are cell culture media and methods culturing host cells expressing polypeptides to reduce the level of trisulfide bonds in polypeptides produced by the host cells. [Continued on next page] WO 2017/196902 A3 MUNRO DOI 01011MMEM31111111111 1111111101101 1111111110fin (84) Designated States (unless otherwise indicated, for every kind of regional protection available): ARIPO (BW, GH, 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, F1, FR, GB, GR, HR, HU, 1E, 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). 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)) (88) Date of publication of the international search report: 22 February 2018 (22.02.2018)