Development and characterization of a potential diagnostic monoclonal antibody against capsid protein VP1 of the chicken anemia virus.
- Author:
Yi Yang LIEN
1
;
Chi Hung HUANG
;
Fang Chun SUN
;
Shyang Chwen SHEU
;
Tsung Chi LU
;
Meng Shiunn LEE
;
Shu Chin HSUEH
;
Hsi Jien CHEN
;
Meng Shiou LEE
Author Information
- Publication Type:Original Article
- Keywords: chicken anemia virus; immunoaffinity column; immunohistochemistry; monoclonal antibody; VP1
- MeSH: Animals; Antibodies, Monoclonal/biosynthesis/genetics/*immunology; Antigens, Viral/analysis; Capsid Proteins/genetics/*immunology; Chicken anemia virus/genetics/*immunology; *Chickens; Circoviridae Infections/blood/immunology/*veterinary/virology; Escherichia coli/genetics; Immunohistochemistry/veterinary; Liver/virology; Mice; Mice, Inbred BALB C; Microscopy, Fluorescence/veterinary; Poultry Diseases/blood/immunology/*virology; Specific Pathogen-Free Organisms; Thymus Gland/virology
- From:Journal of Veterinary Science 2012;13(1):73-79
- CountryRepublic of Korea
- Language:English
- Abstract: Chicken anemia virus (CAV) is an important viral pathogen that causes anemia and severe immunodeficiency syndrome in chickens worldwide. In this study, a potential diagnostic monoclonal antibody against the CAV VP1 protein was developed which can precisely recognize the CAV antigen for diagnostic and virus recovery purposes. The VP1 gene of CAV encoding the N-terminus-deleted VP1 protein, VP1Nd129, was cloned into an Escherichia (E.) coli expression vector. After isopropyl-beta-D-thiogalactopyronoside induction, VP1Nd129 protein was shown to be successfully expressed in the E. coli. By performing an enzyme-linked immunoabsorbent assay using two coating antigens, purified VP1Nd129 and CAV-infected liver tissue lysate, E3 monoclonal antibody (mAb) was found to have higher reactivity against VP1 protein than the other positive clones according to the result of limiting dilution method from 64 clones. Using immunohistochemistry, the presence of the VP1-specific mAb, E3, was confirmed using CAV-infected liver and thymus tissues as positive-infected samples. Additionally, CAV particle purification was also performed using an immunoaffinity column containing E3 mAb. The monoclonal E3 mAb developed in this study will not only be very useful for detecting CAV infection and performing histopathology studies of infected chickens, but may also be used to purify CAV particles in the future.
