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[目的]获得抗Cu2+的单克隆抗体。[方法]以异硫氰酸苄基乙二胺四乙酸(ITCBE)为双功能螯合剂,使Cu2+分别与载体蛋白牛血清白蛋白(BSA)、卵清蛋白(OVA)偶联,获得免疫原(Cu-ITCBE-BSA)和检测原(Cu-ITCBE-OVA)。用非变性聚丙烯凝胶电泳鉴定偶联结果。免疫Balb/C小鼠,通过细胞融合,获得能稳定分泌抗Cu2+的杂交瘤细胞株,并通过亲和层析法纯化腹水获得抗Cu2+的单克隆抗体。[结果]获得了1株能稳定分泌抗Cu2+的单克隆抗体的细胞株(4A6),所分泌的抗体亚类为IgG1,细胞培养上清效价可达1∶1.0×103,腹水效价可达1∶6.4×105。以该细胞株接种Balb/C小鼠腹腔,获得抗Cu2+的腹水型单抗;该腹水经过亲和层析法纯化后,纯度可达95%。[结论]获得了抗Cu2+的单克隆抗体,为环境水样中Cu2+的免疫学检测奠定基础。
[Objective] To obtain anti-Cu2 + monoclonal antibody. [Method] With the ITCBE as bifunctional chelator, the Cu2 + was coupled with the carrier protein BSA and OVA respectively to obtain the immunogen (Cu-ITCBE-BSA) and detection of the original (Cu-ITCBE-OVA). Coupling results were confirmed by non-denaturing polypropylene gel electrophoresis. Balb / C mice were immunized to obtain hybridoma cell lines stably secreting anti-Cu2 + by cell fusion. Anti-Cu2 + monoclonal antibodies were obtained by purification of ascites by affinity chromatography. [Results] A cell strain (4A6) secreting monoclonal antibody against Cu2 + was obtained. The secreted antibody subclass was IgG1. The titer of the cell culture supernatant was 1: 1.0 × 103. The ascites titer was Up to 1: 6.4 × 105. The cell line was inoculated intraperitoneally with Balb / C mice to obtain anti-Cu2 + ascites monoclonal antibody. The purity of ascites was 95% after purification by affinity chromatography. [Conclusion] Monoclonal antibodies against Cu2 + were obtained and laid the foundation for the immunological detection of Cu2 + in environmental water samples.