[从一名黑色素瘤患者构建及筛选人噬菌体抗体库]
[Construction and screening of the human phage antibody library from a patient with melanoma].
作者信息
Yang Jun, Gao Tian-wen, Wang Gang, Wang Lei, Shen Zhu, Li Chun-ying
机构信息
Department of Dermatology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.
出版信息
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2004 Sep;20(5):588-91.
AIM
To construct and screen the human phage antibody library from a patient with melanoma.
METHODS
Total RNA was extracted from the peripheral blood mononuclear cells (PBMCs) of a patient with melanoma who has survived for twenty years. The heavy chain Fd fragments and kappa light chains were amplified by RT-PCR. The amplified products were cloned into phagemid vector to construct Fab antibody library. The library was panned with two kinds of melanoma cells for 4 rounds.
RESULTS
Antibody library contained 1.2 x 10(8) different clones. Specific phage antibodies against one of the two melanoma cells were obtained after 4 rounds of panning.
CONCLUSION
Specific human phage antibodies against melanoma cells were obtained.
目的
构建并筛选来自一名黑色素瘤患者的人噬菌体抗体库。
方法
从一名存活20年的黑色素瘤患者的外周血单个核细胞（PBMCs）中提取总RNA。通过逆转录聚合酶链反应（RT-PCR）扩增重链Fd片段和κ轻链。将扩增产物克隆到噬菌粒载体中构建Fab抗体库。该库用两种黑色素瘤细胞进行4轮淘选。
结果
抗体库包含1.2×10⁸个不同的克隆。经过4轮淘选后获得了针对两种黑色素瘤细胞之一的特异性噬菌体抗体。
结论
获得了针对黑色素瘤细胞的特异性人噬菌体抗体。
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