Recombinant Anti-Botulinum Neurotoxin A Single-Chain Variable Fragment Antibody Generated Using a Phage Display System

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Authors
  1. Mah, D.C.W.
  2. Hu, W-G,
  3. Pon, J.K.
  4. Masri, S.A.
  5. Fulton, R.E.
  6. Monette, P.L.
  7. Nagata, L.P.
Corporate Authors
Defence R&D Canada - Suffield, Ralston ALTA (CAN)
Abstract
A recombinant single-chain fragment variable antibody (scFv) to botulinum A neurotoxin (BoNT/A) was developed. BALB/C mice were immunized with BoNT/A. Splenomic RNA was isolated from the hyperimmune mice and used to prepare a cDNA library, from which the variable regions of the heavy and light chain antibody genes were generated and connected by a DNA linker. The resulting scFv genes were cloned into the phagemid vector pCANTABS in order to construct phage display scFv libraries. Individual anti-BoNT/A phage clones were isolated from the phage display libraries by immunoaffinity selection using immobilized BoNT/A and further evaluated by enzyme-linked immunosorbant assay, immunoprecipitation and Western blotting. Forty-eight clones were found to be BoNTA/A reactive. The most reactive clone, designated D12, was selected for further study. The scFv gene of D12 was subcloned into a Pichia pastoris vector, and expression in yeast was evaluated.
Keywords
Biological strains;Recombinant antibody technology;Botulinum neurotoxin;Recombinant antibody technology;Single chain variable fragment antibody;Phage display technology
Report Number
DRDC-SUFFIELD-SL-2003-031 — Scientific Literature
Date of publication
30 Nov 2003
Number of Pages
8
DSTKIM No
CA023667
CANDIS No
521009
Format(s):
Hardcopy;Document Image stored on Optical Disk

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