Chen L, Wang S, Zhang Y
Institute of Basic Medical Sciences, CAMS, Beijing.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 1997 Apr;19(2):150-3.
For rescueing the heavy chain variable region (VH) gene of CM-1--a human monoclonal antibody against breast cancer, total RNA was extracted from the CM-1 hybridoma cells. After preparing its cDNA by reverse transcription and amplifying by PCR, a DNA fragment with about 400 bp was obtained. With an internal primer, the fragment was directly sequenced and 344 bases which consisted well to known human antibody VH gene construction were read out. In addition, its corresponding amino acid sequences coincided with the 1-3 group canonical structure of human antibody variable region. So we suggested that CM-1 could be considered as an individual of human antibody family, and should aid the preparation of antibody molecules.
为克隆抗人乳腺癌单克隆抗体CM-1重链可变区(VH)基因,从CM-1杂交瘤细胞中提取总RNA。经逆转录制备cDNA并进行PCR扩增后,获得一条约400bp的DNA片段。用内部引物对该片段直接测序,读出344个与已知人抗体VH基因结构完全匹配的碱基。此外,其相应的氨基酸序列与人抗体可变区的1-3组典型结构相符。因此,我们认为CM-1可被视为人类抗体家族的一员,有助于抗体分子的制备。