Development of Functional Second Generation Monoclonal Single Chain Variable Fragment (SCFV) Antibodies Against Venezuelan Equine Encephalitis Virus: Cloning, Expression, Sequencing, and Functional Analysis of Two SCFV Antibodies

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Authors
  1. Alvi, A.Z.
  2. Suresh, M.R.
  3. Stadnyk, L.L.
  4. Nagata, L.P.
  5. Fulton, R.E.
Corporate Authors
Defence R&D Canada - Suffield, Ralston ALTA (CAN);Alberta Univ, Edmonton ALTA (CAN) Faculty of Pharmacy and Pharmaceutical Sciences
Abstract
Monoclonal antibodies (Mabs) against Venezuelan equine encephalitis (VEE) virus has been previously used for diagnostic purposes as well as for immunotherapy in mice. We have generated single chain variable fragment (ScFv) antibodies from four well-characterised Mabs against VEE virus, by cloning variable regions of the heavy (VH) and the light (VL) chain antibody genes, connected by a DNA linker, in phagemid expression vector pCANTAB5E. Mab clones lA4A1 and 5B4D6 were successfully cloned as ScFv in Escherichia coli strain TG-1 and expressed as -30 KDa ScFv proteins which were functional in recognising VEE virus by ELISA. Results were reproduced in Escherichia coli strain HB2151 where the same clones, designated A116 and D66, were expressed minaly as soluble periplasmic proteins. The 30 KDn A116 and D66 ScFv bound VEE virus by ELISA with differential binding specificities. TRUNCATED
Keywords
Alpha viruses;Functional ScFv;Second generation antibodies;Single chain antibodies;Cloning;Expression;Nucleic acid sequencing;Anti E-tag antibodies;Recombinant phage;antibody system (RPAS);Phage display technology;VEE virus;Monoclonal antibodies;Equine encephalomyelitis viruses
Report Number
DRDC-SUFFIELD-TR-1999-033 — Technical Report
Date of publication
01 May 2002
Number of Pages
55
DSTKIM No
CA021647
CANDIS No
518356
Format(s):
Hardcopy;Document Image stored on Optical Disk

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