抗乳腺癌单抗CDI315B可变区基因的序列分析

SEQUENCE OF V ARIABLE VREGION GENES FOR ANTI BREAST CANCER McAb CDI 315B

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 and sequence the V region genes of anti breast cancer McAb CDI 315B Methods:The entire V H and V K genes of anti breast cancer McAb CDI 315B were amplified by RT PCR method from CDI 315B hybridoma cell,using 5′primer and 3′primer as leader sequences for the light chain gene and heavy chain gene,respectively Their nucleotide sequences were determined using Sanger′s method Results:The cDNA sequences of V H and V K region genes were compared to the murine sequences from Kabat database,in which the CDI 315B V K region belongs to the 4/5 group V K subgroup VI and the CDI 315 V H region belongs to V H2 family Conclusion:This result will lay a foundation for the construction of CDI 315B human mouse chimeric light chain and heavy chain