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

Abstract

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.

• Authors: Ying 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
• Journal: Immunology letters (Immunol Lett) Vol. 163 Issue 1 Pg. 8-13 (Jan 2015) ISSN: 1879-0542 [Electronic] Netherlands
• PMID: 25448704 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 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

Topics

• 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)