Identification of the B-cell epitopes on N protein of type 2 porcine reproductive and respiratory syndrome virus, using monoclonal antibodies

International Journal of Biological Macromolecules
Gaiping ZhangAiping Wang

Abstract

Porcine reproductive and respiratory syndrome virus (PRRSV) nucleocapsid protein (N) is the immunodominant region of PRRSV viral proteins. However, B cell epitopes present on N protein have not been well characterized. In this study, The ORF7 gene was amplified by RT-PCR and inserted into the expression vector pET-28a, the constructed pET-28a-N was transformed into Escherichia coli BL21. After expression, purification and identification, the recombined N protein was used as the target to generate monoclonal antibody (mAb). Strains of hybridoma cells secreting anti-N mAbs were obtained by the hybridoma technique. Three of them (named 1G4, 1C6, 3D11) were specifically reacted with PRRSV by IPMA and IFA. To identify the B-cell epitopes within PRRSV N protein, six serial overlapping synthesized peptides (P1-P6) covering the whole region of N were used to define the epitopes recognized by 1G4, 1C6, 3D11. Importantly, after the identification of dot-ELISA and indirect ELISA, we found that 1-15aa was a new epitope which had never been reported before and that it was highly conserved among some HP-PRRSV isolates of type 2 PRRSV. The results of this study might open new perspectives on the detection of PRRSV and have important implicati...Continue Reading

Citations

May 24, 2020·Applied Microbiology and Biotechnology·Min JiangAiping Wang
Jul 30, 2019·International Journal of Biological Macromolecules·Xibao ShiGaiping Zhang
Oct 19, 2019·Comparative Immunology, Microbiology and Infectious Diseases·Jingyuan ZhangSurong Sun

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