SG11201808700QA - Large scale pei-mediated plasmid transfection - Google Patents

Abstract

INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property 11111 1411 0 11101 010 11111 01 100011101110 MIN 11111 00 11110 110 HMI Organization International Bureau (10) International Publication Number (43) International Publication Date .....0\"\" WO 2018/007873 A4 11 January 2018 (11.01.2018) W I PO I PCT — (51) (21) (22) (25) Filing Language: (26) (30) (71) (71) International Patent Classification: C12N 15/86 (2006.01) International Application International Filing Date: Publication Language: Priority Data: 62/322,651 14 April 2016 (14.04.2016) PCT/US2017/025681 04 April 2017 (04.04.2017) Applicant: TRIZELL LTD. Oakley Road, Chinnor OX39 4TW (GB). (72) Inventors: LESCH, Hanna; Microkatu 1 s, FIN-70210 Kuopio (FI). VALKAMA, Anniina; Microkatu 1s, Number: FIN-70210 Kuopio (FI). MALINEN, Joonas; Microkatu PCT/IB2017/001205 ls, FIN-70210 Kuopio (FI). LIPPONEN, Eevi; Microkatu 1s, FIN-70210 Kuopio (FI). LEINONEN, Hanna; Mi- crokatu 1, Kuopio (FI). 25 April 2017 (25.04.2017) (74) Agent: PHARMACEUTICAL PATENT ATTOR- English NEYS, LLC; 55 Madison Ave, 4th Floor, Morristown, NJ 07960-7397 (US). English (81) Designated States (unless otherwise indicated, for every kind of national protection available): AE, AG, AL, AM, US 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, US DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, [GB/GB]; Sanderum House HR, HU, ID, IL, IN, IR, IS, 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, PHARMACEUTICAL PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, LLC [US/US]; 55 Madison SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, NJ 07960-7397 (US). TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. Applicant (for US only): PATENT ATTORNEYS, Ave, 4th Floor, Morristown, (54) ,1 precipitate 0 transfection Title: LARGE SCALE PEI-MEDIATED PLASMID TRANSFECTION pump titer viral vectors of the PEI-plasmid improve PEI-based high A. Recirculation of the medium Medium c mtaining Tra nsfection Mix iCELLis500 Bioreactor = pump a way to = — = — _ = = B. Perfusion of the medium = = fixed Figure 2 : We have found a bed or adherent cell culture which in prior art approaches by modifying how pH pump by using PEI as a transfection adherent — 0- iCELLis500 Bioreactor = Fresh Medium Used me( I i H r 1 = pump agent, We have also found plasmid transfection way to make possible bioreactors clogged and CO2 are managed using PEI to produce while avoiding formation 71' IN en GC IN © c ::: :) (57) -- -- GC in ei large-scale during transfection. bioreactor substrates. C [Continued on next page] WO 2018/007873 A4 MIDEDIMOMOIDEIREEMOOMOVIEDEIIHMEION (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, 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(1)) — as to applicant's entitlement to apply for and be granted a patent (Rule 4.17(11)) — as to the applicant's entitlement to claim the priority of the earlier application (Rule 4.17(iii)) — of inventorship (Rule 4.17(iv)) Published: — with international search report (Art. 21(3)) — with amended claims (Art. 19(1)) — the filing date of the international application is within two months from the date of expiration of the priority period (Rule 26bis.3) Date of publication of the amended claims: 01 March 2018 (01.03.2018)