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目的 :克隆抗乳腺癌 CDI315 B单抗可变区基因 ,并对其进行序列分析。方法 :利用前导肽序列设计引物 ,采用 RT-PCR技术从分泌抗人乳腺癌单克隆抗体的杂交瘤细胞株 CDI315 B分离克隆了抗体轻重链可变区基因 ,用 Sanger双脱氧末端终止法测定扩增的 CDI315 B单抗 VK 和 VH 基因的序列 ,对其进行序列分析。结果 :和国际标准的小鼠 Kabat分类体系进行对比分析 ,CDI315 B单抗的 VK属于第 4或第 5组的第 6亚组 (VI) ;CDI315 B单抗的 VH 则属于第 2族 (VH2 )。结论 :抗乳腺癌 CDI315 B单抗可变区基因的克隆及序列分析为构建人鼠嵌合轻链和人鼠嵌合重链打下基础。
OBJECTIVE: To clone the variable region gene of anti-breast cancer CDI315 B monoclonal antibody and analyze its sequence. Methods: Primer was designed by using the leader peptide sequence. The variable light chain variable region gene was cloned and isolated from the hybridoma cell line CDI315 B secreting anti-human breast cancer monoclonal antibody by RT-PCR. The variable region gene was detected by Sanger dideoxy terminator method The sequence of the increased CDK315B mAb VK and VH genes was sequenced. Results: Compared with the international Kabat classification system, the VK of CDI315 B monoclonal antibody belongs to the sixth subgroup (group VI) of group 4 or group 5. The VH of CDI315 B monoclonal antibody belongs to group 2 (VH2 ). CONCLUSION: Cloning and sequence analysis of variable region genes of anti-breast cancer CDI315 B monoclonal antibody lay the foundation for the construction of human chimeric light chain and human chimeric heavy chain.