Patent SG-11201804400S-A (SG)
Title: Scalable methods for producing recombinant adeno-associated viral (aav) vector in serum-free suspension cell culture system suitable for clinical use
Inventor: QU GUANG
IPC: C12Q1/70
Filed: 2016-12-01
Abstract: INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property -' Organization International Bureau rist0) (43) International Publication Date .... .....r ....) 8 June 2017 (08.06.2017) W I P0 1 PCT ID Hit (10) WO 11111111111111111111111111 International II 2017/096039 III 1111111111 Publication Ell III Number Al IIIIIIIIIItIIIIIIIIIIIIIII (51) International Patent Classification: DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, C12N 5/10 (2006.01) A61K 48/00 (2006.01) HN, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KN, KP, KR, C12N 7/00 (2006.01) A61K 31/7088 (2006.01) KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, C12N 7/02 (2006.01) C12Q 1/70 (2006.01) MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, C12N 7/01 (2006.01) 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, (21) International Application Number: TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, PCT/US2016/064414 ZW. (22) International Filing Date: (84) Designated States (unless otherwise indicated, for every 1 December 2016 (01.12.2016) kind of regional protection available): ARIPO (BW, GH, (25) Filing Language: English GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, (26) Publication Language: English TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, (30) Priority Data: DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, 62/261,815 1 December 2015 (01.12.2015) US 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, (71) Applicant: SPARK THERAPEUTICS, INC. [US/US]; GW, KM, ML, MR, NE, SN, TD, TG). 3737 Market Street, Suite 1300, Philadelphia, Pennsylvania 19104 (US). Declarations under Rule 4.17: as to applicant's entitlement to apply for and be granted a (72) Inventors: QU, Guang; 22 Beaver Dam Drive, Sick- lerville, New Jersey 08081 (US). LU, Lin; 439 Robin Hill patent (Rule 4.170) Road, Wayne, Pennsylvania 19087 WRIGHT, John — as to the applicant's entitlement to claim the of the (US). priority Fraser; 68 River Birch Circle, Princeton, New Jersey earlier application (Rule 4.17(iii)) 08540 (US). — of inventorship (Rule 4.17(iv)) (74) Agent: BEDGOOD, Robert; Pillsbury Winthrop Shaw Published: Pittman LLP, P.O. Box 10500, McLean, Virginia 22102 (US). — with international search report (Art 21(3)) before the expiration of the time limit for amending the (81) Designated States (unless otherwise indicated, for every claims and to be republished in the event of receipt of kind of national available): AE, AG, AL, AM, protection 48.2(h)) AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, amendments (Rule BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, (54) Title: SCALABLE METHODS FOR PRODUCING RECOMBINANT ADENO-ASSOCIATED VIRAL (AAV) VECTOR IN SERUMSUSPENSION CELL CULTURE SYSTEM SUITABLE FOR CLINICAL USE = -FREE No free PEI Add 1/2 free PEI Add 1/3 free PEI , :,,,,* ';' ••;\ .4.!A. , 0 N..;,44.• k:4 1 / 4 c) re:::\t, , ,,` .x., „ ,• ..,:„---, ..t., ‘, ., .\\''''• , th: cr, o it-- 1-1 N (57) : Methods and compositions for transfecting compositions are disclosed in which transfection efficiency significantly increased by contacting the cells being transduced with Q polyethyleneimine (PEI) that is free of nucleic acid during vectors generated according to the disclosed methods and compositions are also disclosed. . is .:.\ FIG. cells the \ A' ‘ • I,VN \ , ' 4* N - \ \ \ - • \•: '' ' ., ':s •c•`,,,, \ s4 ,, 4, = ' , '. \ '''‘.; \ I :'+. ., %. • s , .. \ • ..•, \ . „.„\:.. .- - .:\ --,, , ,„:„ 3A with plasmids are disclosed. In certain embodiments, methods and transfection process. Therapeutically useful adeno-associated viral