Development of a monoclonal antibody against PRRSV glycoprotein 3 using an immuodominant peptide as immunogen.

Abstract

Glycoprotein 3 (GP3), a highly glycosylated membrane protein, is a protective antigen and minor structural protein of porcine reproductive and respiratory syndrome virus (PRRSV), and plays a crucial role in virus assembly and infection. In the present study, we synthesized 23 overlapping peptides span GP3 protein sequence and used pig anti-PRRSV serums to identify immunodominant peptides by indirect ELISA. Five immunodominant peptides GP3-P3, P4, P5, P6 and P7 were identified and GP3-P4 (P55LCPTRQAAAEILEPGKS72) was conjugated to carrier protein BSA. One mAb 1E5 against GP3 was generated from BALB/c mice immunized with the conjugates BSA-P4. The Characterization of mAb was identified by Western blot, Dot-ELISA, IPMA and IFA. We found that mAb 1E5 can specifically react with HP-PRRSV strains but not C-PRRSV or NADC30-like PRRSV strains tested in this study. Site-directed alanine substitution analysis revealed that 8 amino acid residues were involved in antibody binding, among them E65, L67 and P69 were critical residue recognized by mAb 1E5. Taken together, this study provided a novel strategy for generating specific mAbs against virus proteins by using immunodominant peptides as targets, and the mAb 1E5 may be useful for development of rapid differential detection method differentiating HP-PRRSV from C-PRRSV and NADC30-like PRRSV.

• Authors: Chao Liang, Hongliang Liu, Jingming Zhou, Yumei Chen, Peiyang Ding, Xifang Zhu, Mengmeng Wang, Menghao Ding, Aiping Wang
• Journal: International journal of biological macromolecules (Int J Biol Macromol) Vol. 187 Pg. 683-689 (Sep 30 2021) ISSN: 1879-0003 [Electronic] Netherlands
• PMID: 34333004 (Publication Type: Journal Article)
• Copyright: Copyright © 2021. Published by Elsevier B.V.

Chemical References

• Antibodies, Monoclonal
• Antigens, Viral
• Glycoproteins
• Immunodominant Epitopes
• Peptide Fragments
• Viral Proteins

Topics

• Animals
• Antibodies, Monoclonal (immunology)
• Antibody Specificity
• Antigens, Viral (administration & dosage, genetics, immunology)
• Cell Line, Tumor
• Enzyme-Linked Immunosorbent Assay
• Epitope Mapping
• Female
• Glycoproteins (administration & dosage, genetics, immunology)
• Immunization
• Immunodominant Epitopes
• Mice, Inbred BALB C
• Peptide Fragments (administration & dosage, genetics, immunology)
• Porcine respiratory and reproductive syndrome virus (genetics, immunology)
• Sus scrofa
• Viral Proteins (administration & dosage, genetics, immunology)
• Mice