抗RAET1G2单克隆抗体的研制和初步鉴定

Preparation and preliminary identification of monoclonal antibodies against human RAET1G2

目的:制备鼠源抗RAET1G2单克隆抗体,并对其特异性进行鉴定。方法:以原核表达的RAET1G2蛋白为抗原免疫BALB/c小鼠,运用B淋巴细胞杂交瘤技术制备抗RAET1G2单克隆抗体;用免疫双扩散方法鉴定Ig亚类;Western blot鉴定单克隆抗体的特异性。结果:获得了4株可分泌特异性抗RAET1G2抗体的杂交瘤细胞株7A11、9C6、10F9和12F8,Ig亚类均为IgG1。结论:制备的杂交瘤细胞株能稳定分泌特异性的抗RAET1G2抗体,并且所分泌的单克隆抗体都能识别天然的RAET1G蛋白,为进一步研究游离性的RAETlG2分子与肿瘤进展的关系以及分析各种肿瘤细胞表面的RAET1G表达情况创造了条件。

Objective：To prepare and identify the monoclonal antibody against RAET1G2.Methods：Prokaryotic expressed human RAETlG2 was used as an antigen to immunize BALB/c mice, and prepare monoclonal antibodies by means of the B lymphocyte hybridoma technique.Ig subclass and specificity of monoclonal antibodies was analysed by double immunodiffusion and western blot respectively.Results：4 hybridoma cell lines（7A11,9C6,10F9 and 12F8） secreting anti-RAET1G2 mAbs were obtained. Ig subclass of mAbs belonged to IgG1.Conclusion：We obtained four strains of hybridoma cells secreting anti-RAET1G2 antibodies.All of the antibodies could recognize RAET1G expressed on the CHO cell surface.It could be used to study the relation of RAET1G2 with tumor progression and to observe the expression of RAET1G2 on the surface of tumor cells.