Invention Grant
- Patent Title: Means and methods for the determination of the biological activity of BoNT/E in cells
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Application No.: US15536839Application Date: 2015-12-18
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Publication No.: US10900955B2Publication Date: 2021-01-26
- Inventor: Cornelia Bruenn , Gerd Mander
- Applicant: MERZ PHARMA GMBH & CO. KGAA
- Applicant Address: DE Frankfurt am Main
- Assignee: MERZ PHARMA GMBH & CO. KGAA
- Current Assignee: MERZ PHARMA GMBH & CO. KGAA
- Current Assignee Address: DE Frankfurt am Main
- Agency: J.A. Lindeman & Co., PLLC
- Priority: EP14199282 20141219
- International Application: PCT/EP2015/080395 WO 20151218
- International Announcement: WO2016/097243 WO 20160623
- Main IPC: G01N33/573
- IPC: G01N33/573 ; G01N33/50 ; C07K16/18 ; C07K16/12

Abstract:
The present invention pertains to a polyclonal or monoclonal antibody specifically binding to BoNT/E-cleaved SNAP-25. Further, the invention provides a method for directly determining the biological activity of BoNT/E in cells, comprising the steps of: a) incubating cells susceptible to BoNT/E intoxication with BoNT/E for a time and under conditions which allow for the BoNT/E to exert its biological activity; b) fixing the cells and, optionally, permeabilizing the cells with a detergent; c) contacting the cells with at least a first capture antibody specifically binding to non-cleaved and BoNT/E-cleaved SNAP-25, and with at least a second capture antibody specifically binding to BoNT/E-cleaved SNAP-25, wherein the second capture antibody is an antibody of the invention, under conditions which allow for binding of said capture antibodies to the indicated substrates; d) contacting the cells with at least a first detection antibody specifically binding to the first capture antibody, under conditions which allow for binding of said first detection antibody to said first capture antibody, thus forming first detection complexes, and with at least a second detection antibody specifically binding to the second capture antibody, under conditions which allow for binding of said second detection antibody to said second capture antibody, thus forming second detection complexes, and wherein the first detection antibody is different from the second detection antibody; e) determining the amount of the first and second detection complexes of step d); and f) calculating the amount of SNAP-25 cleaved by BoNT/E in said cells by means of the second detection complexes, thereby determining the biological activity of BoNT/E in said cells. Furthermore, the invention relates to a kit for carrying out the method of the invention.
Public/Granted literature
- US20190023771A1 MEANS AND METHODS FOR THE DETERMINATION OF THE BIOLOGICAL ACTIVITY OF BONT/E IN CELLS Public/Granted day:2019-01-24
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