The patent badge is an abbreviated version of the USPTO patent document. The patent badge does contain a link to the full patent document.

The patent badge is an abbreviated version of the USPTO patent document. The patent badge covers the following: Patent number, Date patent was issued, Date patent was filed, Title of the patent, Applicant, Inventor, Assignee, Attorney firm, Primary examiner, Assistant examiner, CPCs, and Abstract. The patent badge does contain a link to the full patent document (in Adobe Acrobat format, aka pdf). To download or print any patent click here.

Date of Patent:
Apr. 28, 2020

Filed:

Nov. 20, 2015
Applicant:

Merz Pharma Gmbh & Co. Kgaa, Frankfurt am Main, DE;

Inventor:

Karl-Heinz Eisele, Frankfurt am Main, DE;

Assignee:

MERZ PHARMA GMBH & CO. KGAA, Frankfurt am Main, DE;

Attorney:
Primary Examiner:
Int. Cl.
CPC ...
C12Q 1/00 (2006.01); G01N 33/68 (2006.01); G01N 33/569 (2006.01); C07K 14/705 (2006.01);
U.S. Cl.
CPC ...
G01N 33/6809 (2013.01); C07K 14/70571 (2013.01); G01N 33/56911 (2013.01); C07K 2319/61 (2013.01);
Abstract

The present invention pertains to a method for determining the biological activity of a neurotoxin, the method comprising the steps of: (a) expressing a fusion protein comprising (i) an anchor protein, (ii) a reporter protein and (iii) a neurotoxin cleavage site intervening the anchor protein and the reporter protein, in neurotoxin-sensitive cells; (b) incubating the neurotoxin-sensitive cells of (a) with a neurotoxin and cultivating the cells under conditions which allow the neurotoxin to exert its biological activity; (c) permeabilizing the neurotoxin-sensitive cells of (b) under conditions which allow the release of the reporter protein but not the release of the anchor protein from the permeabilized neurotoxin-sensitive cells; and (d) quantifying the activity of the reporter protein released from the cells, thereby determining the biological activity of the neurotoxin. In addition, the invention relates to a fusion protein comprising (i) an anchor protein, (ii) a reporter protein, and (iii) a neurotoxin cleavage site intervening the anchor protein and the reporter protein, for determining the biological activity of a neurotoxin, in neurotoxin-sensitive cells. Further encompassed by the present invention is a kit comprising the fusion protein of the invention. Finally, the invention pertains to the use of a fusion protein of the invention for determining the biological activity of a neurotoxin, in neurotoxin-sensitive cells.


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