Patent ID: 8663942
Filing Date: 2014-03-04
Classification: G01N

Abstract:
1. A method for determining the amount of processed neurotoxin polypeptides in a solution comprising processed neurotoxin polypeptides and partially processed and/or unprocessed neurotoxin polypeptides comprising the steps of: a) contacting a first portion of the solution with a first capture antibody which specifically binds to light chains of processed, partially processed and unprocessed neurotoxin polypeptides under conditions which allow for binding of the antibody to the neurotoxin polypeptides, thereby forming a first antibody-complex; b) contacting the first antibody complex—with a detection antibody which specifically binds to heavy chains of the processed, unprocessed and partially processed neurotoxin polypeptides in the antibody complex formed in step a), whereby a first detection complex is formed; c) contacting a second portion of the solution with a second capture antibody which specifically binds to the linker of partially processed or unprocessed neurotoxin polypeptides under conditions which allow for binding of the antibody to the partially processed or unprocessed neurotoxin polypeptides, thereby forming a second antibody-complex; d) contacting the second antibody-complex with the detection antibody which specifically binds to heavy chains of the processed, unprocessed and partially processed neurotoxin polypeptides in the antibody complex formed in step c), whereby a second detection complex is formed, e) determining the amount of the first detection complex formed in step b) and the amount of second detection complex formed in step d) by a determination of the amount of specifically bound detection antibody, and f) calculating the amount of processed neurotoxin polypeptides based on the amounts of the first and second detection complex determined in step e) by subtracting the amount of the second detection complex from the amount of the first detection complex, thereby determining the amount of processed neurotoxin.