从大容量人源噬菌体抗体库中筛选抗黑素瘤特异性抗体  

A specific human scFv against malignant melanoma selected from a large human phage antibody library

作　　者：李淼[1] 陈宇萍[1] 王刚[1] 李春英[1] 殷河慧[1] 田艳丽[1] 高天文[1] 

机构地区：[1]第四军医大学西京医院皮肤科,陕西西安710032

出　　处：《细胞与分子免疫学杂志》2005年第5期602-604,共3页Chinese Journal of Cellular and Molecular Immunology

摘　　要：目的:用黑素瘤(MM)细胞系筛选大容量人源噬菌体抗体库,获得能与MM细胞特异性结合的人源性单链抗体(scFv)。方法:用完整的MM细胞对大容量人源噬菌体抗体库进行4轮“吸附-洗脱-扩增”筛选,并通过细胞ELISA法对随机挑选的克隆进行抗原结合活性测定,得到的阳性克隆进一步做DNA序列测定和免疫组化染色鉴定。结果:从80个随机挑选的克隆中,筛得1株抗MM细胞的抗体,该抗体对人的鳞癌细胞、角质形成细胞、黑素细胞、角蛋白、胰蛋白酶、转铁蛋白及小鼠IgG等细胞或抗原的结合反应均呈阴性。免疫组化染色显示,该抗体与MM细胞系Libr的染色呈阳性,对黑素细胞和痣细胞的染色呈阴性。DNA测序结果表明,该抗体VH属于人IgGVH5亚群,VL属于人Vκ亚型。结论:通过筛选噬菌体抗体库获得到1株抗MM细胞的特异性抗体,为MM的靶向治疗研究奠定了基础。AIM： To obtain the specific human scFv against malignant melanoma （MM） using phage antibody library technology. METHODS： The library was panned with a certain kind of MM cells for 4 rounds. The antigen binding activities of random clones were tested by ELISA in order to select specific antibodies, which were then examined by DNA sequence analysis and immunohistochemistry. RESULTS： In ELISA and immunohistochemical staining, the positive clone selected from the 80 random clones was able to bind one kind of MM cell specifically, while not able to bind other irrelavant cells or antigens, such as human＇s squamous carcinoma cells, keratinocytes, melanocytes, keratin, trypsinase, transferrin and mouse IgG, etc. The antibody＇s VH genes belonged to human IgG VH5 subgroup and the VL belonged to human VK subtype determined by DNA sequence analysis. CONCLUSION： One specific human phage antibody against MM cells was obtained by phage display technique, which may be useful for further development of tumor-targeted therapies.

关 键 词：噬菌体 抗体库 恶性黑素瘤 单链抗体 

分 类 号：R739.5[医药卫生—肿瘤] R392.11[医药卫生—临床医学]