Construction and Characterization of a Novel Recombinant Single-Chain Variable Fragment Antibody Against Western Equine Encephalitis Virus

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Authors
  1. Nagata, L.P.
  2. Long, M.C.
  3. Jager, S.
  4. Mah, W.
  5. Jebailey, L.
  6. Mah, M.A.
Corporate Authors
Defence R&D Canada - Suffield, Ralston ALTA (CAN)
Abstract
A novel recombinant single-chain fragment variable (scFv) antibody against Western equine encephalitis virus (WEE) was constructed and characterized. Using antibody phage display technology, a scFv was generated from the WEE specific hybridoma, 10B5 E7E2. The scFv was fused to a human heavy chain IgG sub 1 constant region (CH1-CH3) and contained an intact 6 His tag and enterokinase recognition site (RS10B5huFc). The RS10B5huFc antibody was expressed in E. coli and purified by affinity chromatography as a 70-kDa protein. The RS10B5huFc antibody was functional in binding to EE antigen in indirect enzyme-linked immunosorbent assays (ELISAs). Furthermore, the RS10B5huFc antibody was purified in proper conformation and formed multimers. The addition of the human heavy chain to the scFv replaced effector functions of the mouse antibody. The Fc domain was capable of binding to protein G and human complement. The above properties of the RS10B5huFc antibody make it an excellent candidate for immunodetection and immunotherapy studies.
Keywords
WEEV (Western Equine Encephalitis Virus);Sequencing;Recombinant antibody technology;Genetic analysis;ELISA (Enzyme Linked Immunosorben)
Report Number
DRES-SL-1999-076 — Reprint
Date of publication
13 May 2003
Number of Pages
14
Reprinted from
Hybridoma, vol 19, no 1, 2000, p 1-13
DSTKIM No
CA022365
CANDIS No
519228
Format(s):
Hardcopy;Document Image stored on Optical Disk

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