Cloning and Identification of Single Chain Variable Fragment Recombinant Antibodies to Botulinum Neurotoxin A

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Authors
  1. Mah, D.C.W.
  2. Pon, J.K.
  3. Masri, S.
  4. Monette, P.L.
  5. Fulton, R.E.
  6. Nagata, L.P.
Corporate Authors
Defence Research Establishment Suffield, Ralston ALTA (CAN);Canada West Biosciences, Calgary ALTA (CAN);Animal Diseases Research Institute, Lethbridge AB (CAN)
Abstract
Balb/C mice were inoculated with botulinum toxoid followed by up to 2.0 x E5 LD50 of botulinum A neurotoxin (BoNT/A). Splenomic RNA was isolated from hyperimmune mice and used to prepare a cDNA library. DNA fragments encoding recombinant single chain fragment variable (scFv) antibodies to BoNT/A were cloned using an M13 phage display system. Individual clones were obtained by immunoaffinity selection using immobilized BoNT/A and screened by enzyme-linked immunosorbant assay (ELISA). Western blots were then used to confirm the reactivity of these clones. One hundred and twenty-two clones were obtained that reacted to the botulinum toxin A used in the initial inoculations. These were then tested by ELISA using highly purified BoNT/A, and forty-eight were found to be reactive.
Keywords
Recombinant antibody technology;Recombinant DNA;Botulinum neurotoxin;Single chain variable fragment antibody;ScFv;Phage display
Report Number
DRES-TR-1999-190 — Technical Report
Date of publication
01 Dec 1999
Number of Pages
41
DSTKIM No
CA000849
CANDIS No
512612
Format(s):
Hardcopy;Document Image stored on Optical Disk

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