Preparation of mouse monoclonal antibodies against bovine TNF-α

Objective To prepare monoclonal antibodies (mAbs) against bovine tumor necrosis factor-alpha (BoTNF-α). Methods Recombinant BoTNF-α with His tag (rHis-BoTNF-α) was expressed in a prokaryotic system as immunogen, and recombinant BoTNF-α with GST tag (rGST-BoTNF-α) was expressed as detection antigen....

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Published inXi bao yu fen zi mian yi xue za zhi Vol. 35; no. 3; p. 271
Main Authors Tan, Yue, Xu, Zhengzhong, Zhu, Zhaocheng, Xia, Aihong, Sun, Lin, Chen, Xiang, Jiao, Xinan
Format Journal Article
LanguageChinese
Published China 01.03.2019
Subjects
Animals
Antibodies, Monoclonal - immunology
Antibody Specificity
Cattle
Enzyme-Linked Immunosorbent Assay
Hybridomas
Mice
Mice, Inbred BALB C
Tumor Necrosis Factor-alpha
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Summary:Objective To prepare monoclonal antibodies (mAbs) against bovine tumor necrosis factor-alpha (BoTNF-α). Methods Recombinant BoTNF-α with His tag (rHis-BoTNF-α) was expressed in a prokaryotic system as immunogen, and recombinant BoTNF-α with GST tag (rGST-BoTNF-α) was expressed as detection antigen. The mAbs were developed by hybridoma cell technology. The reactivity of mAbs with rHis-BoTNF-α and rGST-BoTNF-α was detected by Western blot analysis. Indirect ELISA was used to detect the titer and specificity of mAbs, and the reactivity to commercialized recombinant BoTNF-α (rBoTNF-α) was also evaluated. The reactivity of mAbs against natural antigens was identified by flow cytometry. Results Seven hybridomas stably secreting anti-rBoTNF-α mAbs were successfully obtained. The results showed that all seven mAbs had good reactivity and specificity. Flow cytometry analysis also showed that mAb 4G4 had good reactivity with natural BoTNF-α antigens. Conclusion The anti-rBoTNF-α mAbs are successfully achieved.
ISSN:1007-8738