稀有血型筛选用小鼠单克隆抗体的制备及鉴定

Preparation and identification of murine monoclonal antibodies for the antigen screening of rare donors

目的制备血型单克隆抗体,用于筛选稀有血型。方法用人红细胞作为免疫原免疫6周龄Balb/c小鼠(雌),通过小鼠B淋巴细胞杂交瘤技术制备单克隆抗体,用免疫原红细胞筛选阳性克隆,用已知表型的稀有血型红细胞筛选鉴定抗体特异性。用1 L培养袋扩大培养,以硫酸铵法粗提培养上清。以商品试剂盒检测抗体的免疫球蛋白亚类。检测抗体反应性,摸索在全自动血型仪上进行抗原筛选的使用条件。结果建立了3株单克隆抗体细胞株。抗-GPA/GPB(IgG1,κ)盐水法反应,抗体效价为102 400;抗-EnaTS(IgM,κ)盐水法反应,抗体效价为204 800;抗-K14(IgG1,κ)菠萝酶法反应,抗体效价为20 480。结论利用得到的单抗可在自动血型仪上实施快速高通量抗原筛选,可用于筛选MkMk型、En(a-)、Ko及K:-14型等稀有血型。

Objective To prepare monoclonal antibodies（mAb） against human blood group antigens for the establishment of rare blood bank.Methods Three 6 week-old female Balb/c mice were immunized with human red blood cells（RBCs）.Mouse splenocytes were fused with P3U1 myeloma cells and the positive hybridoma clones were screened with the immunized RBCs.Rare RBCs with known phenotypes were used to screen and identify the specificity of mAbs.The subclasses were identified by monoclonal antibody isotyping kit.Results Three mouse hybridoma cell lines were established by lymphocyte hybridoma technology.The specificity and titers of these mAbs were anti-GPA/GPB of×102 400,anti-EnaTS of×204 800 and anti-K14 of ×20 480,respectively.Conclusion These mAbs can be used to screen rare donors by automatic blood grouping machine.