猪弓形虫重组MIC3蛋白单克隆抗体的制备及其初步应用

doi:10.11843/j.issn.0366-6964.2013.10.016

Abstract

Abstract:

MIC3 has high immunoreactivity, and plays an important role in recognition, adhesion and invasion for Toxoplasma gondii to host cells. To prepare the monoclonal antibody (McAbs) against recombinant MIC3 (rMIC3) of T. gondii, we expressed rMIC3 in Escherichia coli, and purified rMIC3 was immunized to BALB/c mice. B lymphocytes hybridization technique was applied to prepare the aimed McAb. Positive clones were screened with ELISA, and then were subcloned to establish stable cell lines. Ascites were induced to produce the McAbs, and its specificity was identified by Western blot analysis. The gold immunochromatographic lateral flow assay system was developed. The results indicated that two stable hybridoma cell lines (B7 and D3) were obtained. The McAbs subclasses all belong to IgG2a and their ELISA titers of the ascite fluid were 1:640 000 and 1:1920 000, respectively. Western blotting analysis confirmed that the two McAbs all can identify the protein MIC3 from lysed tachyzoites and purified rMIC3. Purified polyclone antibody D3 was labeled with colloidal gold particle, and the minimum detectable limit of tachyzoite was 4×10⁴ CFU·mL^－1. This study laid the foundation for further study on related applications.

CLC Number:

S852.7

BAI Yun, WANG Hai-yan, WANG Zhan-wei, JI Yan, WU Xu-su,LIU Dong-xia, SHAO Guo-qing. Preparation and Preliminary Application of Monoclonal Antibodies against rMIC3 of Toxoplasma gondii[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2013, 44(10): 1622-1628.

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Preparation and Preliminary Application of Monoclonal Antibodies against rMIC3 of Toxoplasma gondii

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