Patent SG-11201903523Y-A (SG)
Title: In vitro and cell based assays for measuring the activity of botulinum neurotoxins
Inventor: DONG MIN
IPC: G01N33/542
Filed: 2017-10-19
Abstract: INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property 111111101111 0 1110101011111 011101 0 01111H111001H1111111011111111111110111111 Organization International Bureau (10) International Publication Number (43) International Publication Date .....0\"\" WO 2018/075783 A2 26 April 2018 (26.04.2018) WIP0 I PCT (51) International Patent Classification: Declarations under Rule 4.17: G01N 33/542 (2006.01) Cl 2N 9/02 (2006.01) C12Q 1/66 (2006.01) — as to applicant's entitlement to apply for and be granted a patent (Rule 4.17(H)) (21) International Application Number: PCT/US2017/057411 Published: — without international search report and to be republished (22) International Filing Date: upon receipt of that report (Rule 48.2(g)) 19 October 2017 (19.10.2017) — with sequence listing part of description (Rule 5.2(a)) (25) Filing Language: English (26) Publication Language: English (30) Priority Data: 62/410,558 20 October 2016 (20.10.2016) US (71) Applicant: PRESIDENT AND FELLOWS OF HAR- VARD COLLEGE [US/US]; 17 Quincy Street, Cam- bridge, Massachusetts 02138 (US). (72) Inventors: DONG, Min; 12 Croft Lane, Weatogue, Con- necticut 06089 (US). YU, Feifan; 98 Dundee Court, Ab- erdeen, New Jersey 07747 (US). Agent: JARRELL, Brenda Herschbach et al.; Choate, Hall & Stewart LLP, Two International Place, Boston, (74) _ _ Massachusetts 02110 (US). = Designated States (unless otherwise indicated, for every (81) = 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. _ Designated States (unless otherwise indicated, for every — (84) _ _ 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). = ei M QC (54) Title: IN VITRO AND CELL BASED ASSAYS FOR MEASURING THE ACTIVITY OF BOTULINUM NEUROTOXINS IN Ir) (57) : Disclosed herein are means for the detection and characterization of neurotoxins such as botulinum neurotoxin (BoNT) I r ---- or tetanus neurotoxin. The present disclosure provides methods for determining potency and activity of neurotoxins in vitro and in vivo. 0 —. Also disclosed are polypeptides comprising N- and C- terminal fragments of a reporter protein that are split by a linker comprising GO a 1-1 neurotoxin cleavage site. Cleavage of the linker by a neurotoxin decreases reporter protein activity, thereby indicating activity of © the neurotoxin. Compositions and kits comprising the disclosed polypeptides, nucleic acids comprising nucleotide sequences encoding f'1 such polypeptides, and cells expressing such polypeptides are also disclosed. 0