胰腺癌单克隆抗体重链可变区基因的扩增

Amplification of the heavy chain variable gene of a monoclonal antibody to pancreatic carcinoma

目的 从分泌抗体的胰腺癌杂交瘤YPC3细胞扩增单抗重链可变区基因。方法 以一步法提取杂交瘤细胞RNA,通过逆转录及聚合酶链式反应,反应产物以琼脂糖凝胶电泳观察。结果 扩增的基因长度为350bp,对照骨髓瘤细胞SP2/0未见扩增。

Objective To amplify the heavy chain variable gene from a hybridoma YPC3 cell line.Methods Total RNA was isolated by a single rapid step of guanidinium extraction. Reverse transcription and polymerase chain reaction were used to amplify the heavy chain variable gene. Biochemical characterization of the gene was showed by gel electrophoresis.Results A strong amplification of a 350 bp DNA segment was found in hybridoma YPC3 while not any amplification in SP2/0 mouse myeloma cells.Conclusion Successful amplification of the heavy chain variable gene makes it possible to generate engineering antibody to pancreatic carcinoma.