摘要
本实验室获得的单克隆抗体（ｍＡｂ）ＬＣ１在体外及体内实验中，能与不同病理类型的肺癌细胞结合，具有作为肺癌病理诊断与鉴别诊断的辅助工具的潜在应用价值。本文应用分子生物学技术，从分泌ＬＣ１的杂交瘤细胞中提取总ＲＮＡ，并经过反转录、ＰＣＲ分别分离得ＶＬ基因及ＶＨ基因，并通过序列测定对ＶＨ基因和ＶＬ基因进行了分析。结果表明：ＬＣ１的ＶＨ基因为３５９ｂｐ，ＶＬ基因为３３７ｂｐ；从推导出的氨基酸序列分析证实，它们均符合小鼠Ｉｇ可变区的氨基酸序列特征。根据Ｋａｂａｔ分类体系，ＬＣ１的ＶＨ基因中的ＶＨ基因片段隶属于抗体重链第５个家族，其ＶＬ基因中的ＶＬ基因片段隶属于抗体轻链的Ｖκ２１家族。
The whole process of cloning and sequencing of the variable regions of mAb LC 1(V H and V L) had been described. The variable regions gene fragments V H and V L of mAb LC 1 were amplified and cloned by RT PCR. The sequences of the gene fragments were determined and analyzed. The results showed that PCR product of V H region consisted of 359 bp encoding about 119 amino acid residues, and PCR product of V L region contained 337 bp encoding about 112 amino acid residues. The amino acid sequences of polypeptide V H and V L were in agreement with the characterization of the amino acid sequences of known mouse antibodies’ variable region. According to Kabat classified method, mAb LC 1 V H gene segment and V L gene segment belong to the mouse Ig heavy chain subgroup V and κ chain subgroup V κ 21 respectively. The success of cloning of the gene fragments of mAb LC 1’s variable regions lay a benefit basis for the construction and expression of single chain Fv or Fab.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
1998年第2期89-92,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
世界实验室1-IVMCD-1计划资助
关键词
单克隆抗体
可变区
基因克隆
序列分析
monoclonal antibody variable region gene cloning sequence analysis
