牛肠道病毒2型单克隆抗体的制备及其应用研究

doi:10.11843/j.issn.0366-6964.2015.11.015

Abstract

Abstract:

The study was aimed to prepare monoclonal antibody against BEV-2 and develop ELISA for the detection of antigen of bovine enterovirus type 2 (BEV-2) based on the monoclonal antibody.The purified BEV-2 VP1⁺ recombinant protein was used to immunize BALB/c mice，of which splenocytes were fused with SP2/0 cells by PEG4000．The selection of positive hybridomas was conducted.Two hybridomas (designated as 1G1 and 5G6) secreting monoclonal antibodies(mAb) against BEV-2 VP1⁺ protein were selected.Based on the monoclonal antibodies against BEV-2，a sandwich ELISA was established to detect BEV-2 through a series of optimization.The sensitivity of the sandwich ELISA was 100 TCID₅₀•0.1 mL^-1，and this method had no cross-reaction with BVDV，IBRV and BLV.The results showed that the sandwich ELISA was specific，sensitive and reproducible.This is the first domestic report of establishment of sandwich ELISA for the detection of BEV-2 based on the preparation of the monoclonal antibodies，and it provided a basis for development of kit for rapid diagnosing BEV-2.

CLC Number:

S852.659.6

GUO Jin-yu， ZHANG He-xiao，WU Dan，GAO Zhi-qiang，ZHANG Ran，ZHANG Le-cui. Preparation and Application of the Monoclonal Antibodies against Bovine Enterovirus Type 2[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(11): 2025-2031.

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Preparation and Application of the Monoclonal Antibodies against Bovine Enterovirus Type 2

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