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

Immunol Lett. 2015 Jan;163(1):8-13. doi: 10.1016/j.imlet.2014.11.011. Epub 2014 Nov 24.

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.

Keywords: Anti-idiotype; PRRSV; Single-chain antibody.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Neutralizing* / genetics
  • Antibodies, Neutralizing* / immunology
  • Antibodies, Neutralizing* / pharmacology
  • Antibodies, Viral* / genetics
  • Antibodies, Viral* / immunology
  • Antibodies, Viral* / pharmacology
  • Antibody Specificity / genetics*
  • Cell Line
  • Mice
  • Porcine Reproductive and Respiratory Syndrome / drug therapy
  • Porcine Reproductive and Respiratory Syndrome / genetics
  • Porcine Reproductive and Respiratory Syndrome / immunology
  • Porcine Reproductive and Respiratory Syndrome / pathology
  • Porcine respiratory and reproductive syndrome virus* / genetics
  • Porcine respiratory and reproductive syndrome virus* / immunology
  • Rabbits
  • Single-Chain Antibodies* / genetics
  • Single-Chain Antibodies* / immunology
  • Single-Chain Antibodies* / pharmacology
  • Swine
  • Viral Envelope Proteins* / genetics
  • Viral Envelope Proteins* / immunology

Substances

  • Antibodies, Neutralizing
  • Antibodies, Viral
  • Single-Chain Antibodies
  • Viral Envelope Proteins
  • glycoprotein 5, PRRSV