1. Utilization of phage display to identify antigenic regions in the PCV2 capsid protein for the evaluation of serological responses in mice and pigs.
- Author
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Santos MR, Assao VS, Santos FAA, Salgado RL, Carneiro AP, Fietto JLR, Bressan GC, de Almeida MR, Lobato ZIP, Ueira-Veira C, Goulart LR, and Silva-Júnior A
- Subjects
- Animals, Antibodies, Neutralizing biosynthesis, Antibodies, Neutralizing blood, Antibodies, Neutralizing immunology, Antibodies, Viral biosynthesis, Antibodies, Viral blood, Capsid Proteins chemistry, Circoviridae Infections immunology, Circoviridae Infections prevention & control, Circoviridae Infections virology, Circovirus chemistry, Circovirus genetics, Enzyme-Linked Immunosorbent Assay, Epitopes chemistry, Epitopes immunology, Epitopes isolation & purification, Mice, Neutralization Tests, Peptides chemistry, Peptides isolation & purification, Sensitivity and Specificity, Swine, Swine Diseases blood, Swine Diseases immunology, Swine Diseases virology, Viral Vaccines immunology, Antibodies, Viral immunology, Capsid Proteins immunology, Cell Surface Display Techniques, Circovirus immunology, Peptides immunology
- Abstract
Porcine circovirus 2 (PCV2) is associated with a series of swine diseases. There is a great interest in improving our understanding of the immunology of PCV2, especially the properties of the viral capsid protein Cap-PCV2 and how they relate to the immunogenicity of the virus and the subsequent development of vaccines. Phage display screening has been widely used to study binding affinities for target proteins. The aim of this study was to use phage display screening to identify antigenic peptides in the PCV2 capsid protein. After the selection of peptides, five of them presented similarity to sequences found in cap-PCV2, and four peptides were synthesized and used for immunization in mice: 51-CTFGYTIKRTVT-62 (PS14), 127-CDNFVTKATALTY-138 (PS34), 164-CKPVLDSTIDY-173 (PC12), and 79-CFLPPGGGSNT-88 (PF1). Inoculation with the PC12 peptide led to the highest production of antibodies. Furthermore, we used the PC12 peptide as an antigen to examine the humoral response of swine serum by ELISA. The sensitivity and specificity of this assay was 88.9% and 92.85%, respectively. Altogether, characterization of immunogenic epitopes in the capsid protein of PCV2 may contribute to the improvement of vaccines and diagnostics.
- Published
- 2018
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