IMMUNOLOGICAL ANALYSIS OF THE ACTIVITY OF BOTULINIC TOXIN SEROTYPE A

摘要

1. A composition comprising a carrier, a flexible linker and an SNAP-25 antigen, wherein the flexible linker contains at least three amino acids and the SNAP-25 antigen contains SEQ ID NO: 148. ! 2. The composition of claim 2, wherein the amino acid sequence of the flexible linker and SNAP-25 antigen is SEQ ID NO: 38. ! 3. The selected antibody α-SNAP-25,! where the isolated α-SNAP-25 antibody binds to an epitope containing the C-terminal glutamine of the cleavable bond of the BoNT / A cleavage site of the SNAP-25 cleavage product; ! where the α-SNAP-25 antibody has an association rate constant for an epitope that does not contain a C-terminal glutamine cleavable bond of the BoNT / A cleavage site of the SNAP-25 cleavage product of less than 1 · 101 M-1 s1; and! where the α-SNAP-25 antibody has an equilibrium dissociation constant for the epitope of less than 0.450 nM. ! 4. The isolated α-SNAP-25 antibody of claim 3, wherein the epitope is SEQ ID NO: 32. ! 5. An isolated α-SNAP-25 antibody comprising:! but. the variable region of the heavy chain containing SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 76, SEQ ID NO: 80 or SEQ ID NO: 82; and! b. the variable region of the light chain containing SEQ ID NO: 84, SEQ ID NO: 86, SEQ ID NO: 88, SEQ ID NO: 90 or SEQ ID NO: 92. ! 6. An isolated α-SNAP-25 antibody comprising:! but. the variable region of the heavy chain containing SEQ ID NO: 93, SEQ ID NO: 121 or SEQ ID NO: 100; and! b. the variable region of the light chain containing SEQ ID NO: 105, SEQ ID NO: 110 or SEQ ID NO: 115. ! 7. An isolated α-SNAP-25 antibody containing at least VH CDR3 from SEQ ID NO: 100, VH CDR3 from SEQ ID NO: 101, or VH CDR3 from SEQ ID NO: 102. ! 8. A method for detecting BoNT / A activity, comprising the steps of:! but. treating a cell from a stable cell line with a sample containing BoNT / A, where a cell from a stable cell line is sensitive to BoNT / A intoxication at a concentration of BoNT / A of about 500 pM or less