摘要
目的 :构建噬菌体抗体库并筛选抗黑素瘤细胞的人抗体。方法 :从 1例长期存活的黑素瘤患者的外周血中分离单个核细胞 (PBMC) ,提取总RNA ,并逆转录为cDNA。用PCR扩增多样性κ链基因和重链Fd段基因 ,分别构建κ链库和重链库 ,组合为Fab段噬菌体抗体库 ,用两种黑素瘤细胞系对抗体库进行筛选和鉴定。结果 :所构建的噬菌体抗体库的库容量达 1.2× 10 8,用两种黑素瘤细胞系进行亲和吸附筛选 ,获得针对其中 1种黑素瘤细胞系的特异性抗体。结论 :采用噬菌体抗体库技术 ,成功地从 1例长期存活的黑素瘤患者体内克隆到特异性抗黑素瘤细胞的抗体 。
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 κ 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 1.2×10 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.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2004年第5期588-591,共4页
Chinese Journal of Cellular and Molecular Immunology
