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Monoclonal antibodies directed against VP7 protein of human group B rotavirus

机译:针对人类B型轮状病毒VP7蛋白的单克隆抗体

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The aim of this study was to prepare and identify a monoclonal antibody that binds the viral proteins 7 (VP7 protein) of human group B rotavirus (GBRV) and to describe its immunologic characterization. Human group B rotavirus vp7 gene was successfully ligated into pGEX-KG vector and transformed into Escherichia coli TOP10 cells. The glutathione S-transferases (GST)-fusion protein GST-VP7 was induced by Isopropyl β-D-1-thiogalactopyranoside (IPTG) and immediately purified to immunize BALB/c mice. Splenocytes were then prepared from the immunized mouse and fused with SP2/0 myeloma cell line. In the end we obtained one positive hybridoma cell line stably secreting monoclonal antibody against GST-VP7 protein by indirect enzyme-linked immunosorbent assay (ELISA) and limiting dilution. The production of the monoclonal antibody against GBRV will benefit the further study of GBRV's structures and functions and also lay a solid foundation for the research of disease prevention, clinical diagnosis, and treatment.
机译:这项研究的目的是制备和鉴定与人类B型轮状病毒(GBRV)的病毒蛋白7(VP7蛋白)结合的单克隆抗体,并描述其免疫学特性。将人类B型轮状病毒vp7基因成功连接到pGEX-KG载体中,并转化到大肠杆菌TOP10细胞中。异丙基β-D-1-硫代吡喃半乳糖吡喃糖苷(IPTG)诱导谷胱甘肽S-转移酶(GST)融合蛋白GST-VP7,并立即纯化以免疫BALB / c小鼠。然后从经免疫的小鼠制备脾细胞,并与SP2 / 0骨髓瘤细胞系融合。最后,我们通过间接酶联免疫吸附测定(ELISA)和有限稀释获得了一种稳定分泌抗GST-VP7蛋白单克隆抗体的阳性杂交瘤细胞。抗GBRV的单克隆抗体的产生将有利于GBRV的结构和功能的进一步研究,也为疾病预防,临床诊断和治疗的研究奠定了坚实的基础。

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