肝癌单抗重链可变区基因的克隆及序列测定

Cloning and sequencing heavy chain variable region gene of a monoclonal antibody to hepatocellular carcinoma

从分泌与肝细胞肝癌特异性强并具有良好人体内导向作用的鼠源性单克隆抗体的杂交瘤细胞ＨＡｂ２５中提取ＲＮＡ，逆转录成ｃＤＮＡ，用合成的寡核甘酸引物从中扩增出抗体重链可变区基因。将此基因克隆到ｐＵＣ１９质粒中，测定了此可变区基因的全序列。经计算机分析，结果表明基因长度为３６０ｂｐ，编码１２０个氨基酸，是具有功能性的抗体可变区基因，在核苷酸序列上与小鼠重链ＶＨ１８６．２家族同源性最高。

The isolation of rearranged immunoglobolin genes from a hybridoma cell line,which is prequisite for the construction of a recombinant antibody,can easily be achieved by polymerase chain reaction.Murine McAb HAb25 possesses high specificity for human hepatocellular carcinomain biopsy tissues and human body,minimally reacted with normal tissues,making it a logical candidate for clinical use.We have determined the nucleotide sequence of the variable region of heavy chain of HAb25.Analysis of the nucleotide sequence of the heavy chain reveraled that the VH sequence was homological to the VH186.2 germline gene family,belonging to the mouse heavy chain subgroup Ⅲ.