单克隆抗体
分子生物学
重组DNA
效价
抗体
细胞融合
转染
病毒学
融合蛋白
子类
细胞培养
脾脏
生物
嵌合体(遗传学)
抗原
化学
病毒
免疫学
生物化学
基因
遗传学
作者
Ying-Hao Chu,Xiangdan Gong,Jianwei Gao,Airong Su,Deyan Chen,Hongyong Song,Xi-Lin Wu,Zhiwei Wu
出处
期刊:PubMed
日期:2017-04-01
卷期号:33 (4): 540-545
摘要
Objective To express the V1/V2 domain of HIV-1 subtype CRF01_AE gp120 in eukaryotic cells, and then prepare its monoclonal antibody (mAb) and identify its antigen reactivity. Methods Eukaryotic expression vector of pTriEx-3-V1/V2CNE55 was constructed and transiently transfected into HEK293F cell line. The V1/V2-His chimera protein was purified and injected into BALB/c mice. After the fusion between spleen cells of immunized BALB/c mice and myeloma cells SP 2/0, ELISA was used for screening the positive hybridoma cell clones against the V1/V2 recombinant protein. The specificity, titer and type of its mAb were characterized. Results We obtained a stable hybridoma cell line which secreted anti-HIV-1 AE subtype gp120 V1/V2 domain mAb. The ascite titer of the mAb was 1:81 000, and the type of the mAb was IgG1/κ. Western blotting showed that the mAb could recognize recombinant HIV-1 gp120 of different HIV-1 subtypes. Conclusion The study prepared successfully the anti-HIV-1 V1/V2 domain mAb.
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