Single-chain anti-idiotypic antibody retains its specificity to porcine reproductive and respiratory syndrome virus GP5.

Monoclonal anti-idiotypic antibody (Mab2-5G2) raised against idiotypic antibodies to membrane glycoprotein GP5 of porcine reproductive and respiratory syndrome virus (PRRSV). The variable regions of the heavy chain (VH) and light chain (VL) of Mab2-5G2 were cloned and connected with a (Gly4Ser)3 linker. The recombinant scFv gene was cloned into the pEasy-E1 vector and expressed in E. coli as inclusion bodies. The expressed scFv-His proteins renatured in a pH and urea gradient buffer retained the same immunological properties as that of Mab2-5G2. Renatured scFv-His protein retained the same characteristics as that of Mab2-5G2 by recognizing and binding to Marc-145 cells. Furthermore, renatured scFv-His along with Mab2-5G2 were used to immunize rabbits to produce anti-anti-idiotypic antibodies (Ab3) that neutralized PRRSV infection of Marc-145 cells. These results demonstrated that the expressed scFv-His protein possessed the same characteristics of Mab2-5G2 and will be suitable for future investigations of Mab2-5G2 antibody structure and its ability to interact with potential PRRSV cellular receptor as well as immunological properties against PRRSV infection.
AuthorsYing Yu, Gang Wang, Qiongyi Li, Yongkun Du, Taofeng Du, Yang Mu, Shuqi Xiao, Qin Zhao, Chengbao Wang, Yani Sun, Xingang Xu, Gaiping Zhang, Walter H Hsu, Xuehui Cai, En-Min Zhou
JournalImmunology letters (Immunol Lett) Vol. 163 Issue 1 Pg. 8-13 (Jan 2015) ISSN: 1879-0542 [Electronic] Netherlands
PMID25448704 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
CopyrightCopyright © 2014 European Federation of Immunological Societies. Published by Elsevier B.V. All rights reserved.
Chemical References
  • Antibodies, Neutralizing
  • Antibodies, Viral
  • Single-Chain Antibodies
  • Viral Envelope Proteins
  • glycoprotein 5, PRRSV
  • Animals
  • Antibodies, Neutralizing (genetics, immunology, pharmacology)
  • Antibodies, Viral (genetics, immunology, pharmacology)
  • Antibody Specificity (genetics)
  • Cell Line
  • Mice
  • Porcine Reproductive and Respiratory Syndrome (drug therapy, genetics, immunology, pathology)
  • Porcine respiratory and reproductive syndrome virus (genetics, immunology)
  • Rabbits
  • Single-Chain Antibodies (genetics, immunology, pharmacology)
  • Swine
  • Viral Envelope Proteins (genetics, immunology)

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