芝麻过敏原油质蛋白O leosins单克隆抗体的制备与鉴定

利用细胞融合技术制备小鼠抗油质蛋白杂交瘤细胞，通过间接ELISA方法筛选稳定分泌特异性抗体的杂交瘤细胞株。获得10株杂交瘤细胞，分别命名为1B2、2D8、2F3、3A4、3D7、3F8、4A10、4F12、6A12、6E7，利用Ig亚类鉴定试剂盒鉴定各单克隆抗体的Ig亚型，除1B2、3D7、3F8为IgG1类外其余均为IgG2a类型。将杂交瘤细胞株注射入小鼠腹腔获取腹水，应用Protein A亲和层析法对腹水进行纯化。单抗效价有9株在2.0×105以上。ELISA和Western blotting分析表明这些单抗均能特异性识别芝麻过敏原油质蛋白。

Preparation and Identification of Monoclonal Antibodies against O leosins

To prepare monoclonal antibodies against allergen O leosins from sesame seeds, BALB/c mice were immunized and the splenocytes of the immunized mice were fused with NS-1 myeloma cells by hybridoma technique. Ten hybridoma cell lines secreting McAbs against O leosins were obtained and classified as 1B2, 2D8, 2F3, 3A4, 3D7, 3F8, 4A10, 4F12, 6A12, 6E7, 1B2, 3D7 and 3F8 were subtybes of IgG1 while 2D8, 2F3, 3A4, 4A10, 4F12, 6A12, 6E7 belonged to the subtype of IgG2a. The antibody titers were more than 2.0 × 105. The McAbs were then purified using affinity chromatography on immobilized Protein A and identified by their specificities, subtypes and titers with ELISA and Western blotting. The ten McAbs all recognized recombinant O leosins of sesame seeds. These monoclonal antibodies against O leosins were prepared successfully, which will facilitate to establish the detection of allergens in sesame seeds.