抗Gly m Bd 28K单克隆抗体的制备及其免疫学特性

利用Gly m Bd 28K天然蛋白免疫BALB/c雌性小鼠，取小鼠脾脏细胞与杂交瘤细胞融合，筛选出2株能稳定分泌抗Gly m Bd 28K单克隆抗体的杂交瘤细胞株1E3和2F4。体内诱生腹水法大量制备单克隆抗体，经纯化后，得到2株均为IgG1型的单克隆抗体mAb1E3和mAb2F4。间接ELISA方法测得mAb1E3的效价达到1∶4 10×105，对Gly m Bd 28K蛋白的半数抑制率（IC50）为23.99 μg·L-1，与花生蛋白、核桃蛋白等的交叉反应率均低于0.1%。该单克隆抗体的制备为Gly m Bd 28K致敏蛋白的检测以及抗原表位的鉴定奠定了基础。

Preparation of monoclonal antibodies against Gly m Bd 28K and identification of their immunological properties

To establish hybridoma cell lines that secreted anti Gly m Bd 28K monoclonal antibodies， Gly m Bd 28K protein was used to immunize BALB/c mouse， whose spleen was removed for cell fusion. Two hybridoma cell lines that stably secreted anti Gly m Bd 28K monoclonal antibodies were screened and then were injected into BALB/c mice to prepare monoclonal antibodies. After purification of monoclonal antibody， two strains of cell lines were identified for IgG1， named as mAb1E3 and mAb2F4. Indirect ELISA was used to detect the immunological properties of mAb1E3. The titer of mAb1E3 was 1∶4 10×105， and the IC50 of mAb1E3 to Gly m Bd 28K was 23.99 μg·L-1. The ratios of cross reactivity of mAb1E3 with peanut protein， sesame protein， walnut protein and wheat glutenin were less than 0.1%. We successfully prepared the anti Gly m Bd 28K monoclonal antibodies possessed high sensitivity and specificity. This study provided a basis for the detection of Gly m Bd 28K allergen and the identification of epitopes.