胶质瘤单抗SZ-38可变区基因的克隆和序列分析

Cloning and sequence analysis of variable region genes of anti human glioma monoclonal antibody SZ 38

目的 克隆高特性、高亲和力的胶质瘤靶向载体基因，为制备重组免疫毒素奠定基础。方法 从胶质瘤单抗杂交瘤细胞 Ｓ Ｚ３８ 提取总 Ｒ Ｎ Ａ， Ｒ Ｔ Ｐ Ｃ Ｒ 扩增抗体的重链及轻链可变区基因，并测序分析。结果 重链可变区全长３０６ ｂｐ ，编码１０２ 个氨基酸；轻链可变区全长３１２ ｂｐ ，编码１０４ 个氨基酸，分属重链Ⅲ（ Ｄ） 亚组和 Ｋａｐｐａ 链Ⅵ亚组。结论 新克隆的可变区基因符合功能重排的鼠抗体可变区基因特征，可用于重组免疫毒素的制备。

Objective\ To clone and analyze the variable region genes from an anti human glioma mouse monoclonal antibody hybridoma cell line SZ 38. Methods\ Total RNA was extracted from SZ 38 cells which displayed a high affinity for human glioma. The variable light and heavy chain region (VL and VH) genes were amplified with RT PCR and cloned into PGEM T vector, respectively. Then the two genes were sequenced and analyzed by computer.Results\ The variable region genes contained sequences of CDRs and FRs. The total length of VH gene was 306 bp, encoding 102 amino acids, and that of VL gene was 312 bp, encoding 104 amino acids, which belonged to heavy chain Ⅲ(D) subgroup and kappa chain VI subgroup respectively. Conclusion\ The VH and VL genes were full length and potentially functional, and confirmed as functionally rearranged mouse immunoglobulin variable genes.\;