Back to results
US 2017/0248602 A1Published application
Immuno-based retargeted endopeptidase activity assays
AssigneeAllergan, Inc.
InventorsJoanne Wang, Hong Zhu, Dianne D. Hodges, Ester Fernandez-Salas
Life of this patent
Estimating the term…
Abstract
The present specification discloses a retargeted endopeptidase pharmaceutical wherein the activity has been determined by the methods disclosed.
Claims13
A method of detecting retargeted endopeptidase activity, the method comprising the steps of:
- a. treating a cell from an established cell line with a sample comprising a retargeted endopeptidase, wherein the cell from an established cell line is susceptible to retargeted endopeptidase activity by a retargeted endopeptidase;
- b. isolating from the treated cell a SNAP-25 component comprising a SNAP-25 cleavage product having a carboxyl-terminus at the P1 residue of the BoNT/A cleavage site scissile bond;
- c. contacting the SNAP-25 component with an α-SNAP-25 antibody linked to a solid phase support,
- wherein the α-SNAP-25 antibody binds an epitope comprising a carboxyl-terminus at the P1 residue of the BoNT/A cleavage site scissile bond from a SNAP-25 cleavage product,
- wherein the α-SNAP-25 antibody has a heavy chain variable region comprising an amino acid sequence encoded by the nucleic acid sequence SEQ ID NO:
- 71, SEQ ID NO:
- 75, SEQ ID NO:
- 77, SEQ ID NO:
- 79, or SEQ ID NO:
- 81, or a nucleic acid sequence that is at least 90% identical to SEQ ID NO:
- 71, SEQ ID NO:
- 75, SEQ ID NO:
- 77, SEQ ID NO:
- 79, or SEQ ID NO:
- 81;
- wherein the α-SNAP-25 antibody has a light chain variable region comprising an amino acid sequence encoded by the nucleic acid sequence SEQ ID NO:
- 83, SEQ ID NO:
- 87, SEQ ID NO:
- 89, or SEQ ID NO:
- 91, or a nucleic acid sequence that is at least 90% identical to SEQ ID NO:
- 83, SEQ ID NO:
- 87, SEQ ID NO:
- 89, or SEQ ID NO:
- 91;
- and
- wherein the α-SNAP-25 antibody has a heavy chain variable region comprising the amino acid sequence selected from the group consisting of SEQ ID NO:
- 72, SEQ ID NO:
- 76, SEQ ID NO:
- 78, SEQ ID NO:
- 80, and SEQ ID NO:
- 82;
- and a light chain variable region comprising the amino acid sequence selected from the group consisting of SEQ ID NO:
- 84, SEQ ID NO:
- 88, SEQ ID NO:
- 90, and SEQ ID NO:
- 92;
- d. detecting the presence of an antibody-antigen complex comprising the α-SNAP-25 antibody and the SNAP-25 cleavage product;
- wherein detection by the antibody-antigen complex is indicative of retargeted endopeptidase activity.
35 more elements
An α-SNAP-25 antibody which binds an epitope comprising a carboxyl-terminus at the P1 residue from the BoNT/A cleavage site scissile bond from a SNAP-25 cleavage product;
- wherein the α-SNAP-25 antibody has a heavy chain variable region comprising an amino acid sequence encoded by the nucleic acid sequence SEQ ID NO:
- 71, SEQ ID NO:
- 75, SEQ ID NO:
- 77, SEQ ID NO:
- 79, or SEQ ID NO:
- 81, or a nucleic acid sequence that is at least 90% identical to SEQ ID NO:
- 71, SEQ ID NO:
- 75, SEQ ID NO:
- 77, SEQ ID NO:
- 79, or SEQ ID NO:
- 81;
- wherein the α-SNAP-25 antibody has a light chain variable region comprising an amino acid sequence encoded by the nucleic acid sequence SEQ ID NO:
- 83, SEQ ID NO:
- 87, SEQ ID NO:
- 89, or SEQ ID NO:
- 91, or a nucleic acid sequence that is at least 90% identical to SEQ ID NO:
- 83, SEQ ID NO:
- 87, SEQ ID NO:
- 89, or SEQ ID NO:
- 91;
- and
- wherein the α-SNAP-25 antibody has a heavy chain variable region comprising the amino acid sequence selected from the group consisting of SEQ ID NO:
- 72, SEQ ID NO:
- 76, SEQ ID NO:
- 78, SEQ ID NO:
- 80, and SEQ ID NO:
- 82;
- and a light chain variable region comprising the amino acid sequence selected from the group consisting of SEQ ID NO:
- 84, SEQ ID NO:
- 88, SEQ ID NO:
- 90, and SEQ ID NO:
- 92.
29 more elements
Dependencies are inferred from the claim text.