人源乳腺癌单链抗体基因的构建

Construction of human single-chain antibody gene for breast cancer

目的:构建人源抗乳腺癌单链抗体(scfv)基因片断。方法:首先利用乳腺癌患者癌旁淋巴组织提取总RNA,然后通过RT-PCR分别扩增VH和VL基因片断,再将经纯化的VH和VL基因片断通过“重叠-延伸-拼接PCR”(SOE-PCR)而形成单链抗体(scfv)基因片断。结果:构建了750bp的人源抗乳腺癌单链抗体(scfv)基因片断。结论:成功地构建了人源抗乳腺癌单链抗体基因片断,为进一步构建人源抗乳腺癌单链抗体库提供了试验基础。

Objiective：To construct human single-chain antibody gene fragment for breast cancer.Methods.Total RNA was isolated from tumor adjacent lymphatic tissue of breast cancer patients. Then heavy and light chain （VH and VL） genes of antibody were amplified separately by RT-PCR, and assembled into ScFv genes by SOE-PCR.Results：The human single-chain antibody gene fragment for breast cancer was constructed and its size was about 750bp .Conclusion.The human single-chaln antibody gene fragments were constructed successfully ,which provides a way for constructing the human single-chain antibody library of anti-breast cancer.