Preparation of Monoclonal Antibodies against the Capsid Protein and Development of an Epitope-Blocking Enzyme-Linked Immunosorbent Assay for Detection of the Antibody against Porcine Circovirus 3

Author:

Wang Junli1ORCID,Lei Baishi1,Zhang Wuchao1,Li Lijie1,Ji Jiashuang1,Liu Mandi1,Zhao Kuan12,Yuan Wanzhe123

Affiliation:

1. College of Veterinary Medicine, Hebei Agricultural University, Baoding 071000, China

2. Hebei Veterinary Biotechnology Innovation Center, Hebei Agricultural University, Baoding 071000, China

3. North China Research Center of Animal Epidemic Pathogen Biology, China Agriculture Ministry, Baoding 071000, China

Abstract

Porcine circovirus type 3 (PCV3) is endemic in swine worldwide and causes reproductive disorders, dermatitis and nephrotic syndrome, and multi-organ inflammation. Currently, there is a growing need for rapid and accurate diagnostic methods in disease monitoring. In this study, four monoclonal antibodies (mAbs) against PCV3 capsid proteins were prepared (mAbs 2F6, 2G8, 6E2, and 7E3). MAb 7E3, which had the highest binding affinity for the Cap protein, was chosen for further investigation. A novel B cell epitope 110DLDGAW115 was identified using mAb 7E3. An epitope-blocking (EB) enzyme-linked immunosorbent assay (ELISA) was successfully developed using horseradish-peroxidase-labeled mAb 7E3 to detect PCV3 antibodies in porcine sera. Moreover, the EB-ELISA showed no specific reaction with other porcine disease sera, and the cut-off value was defined as 35%. Compared with the commercial ELISA, the percentage agreement was 95.59%. Overall, we have developed a novel EB-ELISA method that accurately and conveniently detects PCV3 in serum, making it a valuable tool for the clinical detection of PCV3 infection.

Funder

Open Fund project of Fujian University Key Laboratory

Science and Technology Development Projects of Central Guidance for Local

Agricultural Science and Technology Projects of Shijiazhuang

Publisher

MDPI AG

Subject

General Veterinary,Animal Science and Zoology

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