摘要
目的：获取抗人原发性肝癌单克隆抗体重、轻链可变区基因。方法：从分泌高亲和力的抗人原发性肝癌单克隆抗体的杂交瘤细胞株ＳＭ８．１１中提取细胞的总ＲＮＡ，以随机引物反转录合成ｃＤＮＡ，以特异引物进行ＰＣＲ，扩增了单抗的重链可变区（ＶＨ）及轻链可变区（ＶＬ）基因，并进行了序列分析。结果：ＶＨ基因片段长度为３５１ｂｐ，编码１１７个氨基酸残基；ＶＬ基因片段长度为３２４ｂｐ，编码１０７个氨基酸残基。两者均有４个框架区（ＦＲｓ）及３个抗原互补决定区（ＣＤＲｓ），并含有抗体特征性的两个半胱氨酸残基。
Objective: To acquire the variable region genes of heavy and light chain of monoclonal antibody against human primary hepatoma. Methods: Total RNA extracted from hybridoma cell SM8.11 secreting specific McAb against human primary hepatoma was transcripted reversely into cDNA with random primers. The SM8.11 variable region of the heavy chain (VH) and variable region of the light chain (VL) gene fragments were amplified using PCR method and sequenced using the dideoxy chain termination technique. Results: The VH gene consisted of 351 base pairs and encoded 117 amino acids residues; the VL gene consisted of 324 base pairs and encoded 107 amino acids residues. There were four FRs, three CDRs and two characteristic cysteine residues within VH and VL genes, respectively. Conclusion: The success of cloning of McAb SM8.11 VH and VL gene lays a good basis for construction of a recombinant antibody.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
1998年第2期110-112,共3页
Academic Journal of Second Military Medical University
关键词
肝肿瘤
可变区
序列分析
单克隆抗体
hepatoma
antibodies, monoclonal
variable region
gene cloning
sequencing
