| 利用人源化噬菌体抗体库筛选骨唾液酸蛋白的单链抗体 |
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| 引用本文： | 王江涛,王捷. 利用人源化噬菌体抗体库筛选骨唾液酸蛋白的单链抗体[J]. 细胞与分子免疫学杂志, 2011, 27(9): 979-981 |
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| 作者姓名： | 王江涛 王捷 |
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| 作者单位： | 广州军区广州总医院医学实验科,广东广州,510010 |
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| 基金项目： | 2009年广东省卫生厅医学科研基金资助项目(B2009202) |
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| 摘 要： | 目的:从人源化噬菌体抗体库中筛选高亲和、特异结合人骨唾液酸蛋白(BsP)的人源可变区单链抗体(scFv).方法:采用噬菌体表面展示系统,以重组的BSP蛋白为包被抗原,从噬菌体可变区scFv中筛选特异性结合BSP的小分子scFv,经过3轮亲和富集筛选后,再用ELISA方法进一步鉴定与抗原BSP有特异结合活性的scFv阳性克隆,PCR扩增鉴定阳性克隆的插入轻、重链基因片段,并对阳性scFv分子测序和序列分析.结果:筛选得到的scFv片段可以特异地结合BSP蛋白,PCR扩增都得到了长为368 bp、527 bp和935bp的轻链、重链和轻链-连接片段-重链的基因片段,测序结果分析发现上述scFv片段在轻链有11处的氨基酸组成不同,在重链区域氨基酸组成则有3处不同.基因序列分析结果表明符合人源单链可变区抗体基因序列的结构特征.结论:利用噬菌体抗体库技术成功获得BSP蛋白的特异性人源单链可变区抗体.
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| 关 键 词： | 骨唾液酸蛋白 单链抗体 噬菌体文库 |
Screening of single-chain variable fragment (scFv) to bone sialoprotein |
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| Wang Jiang-Tao,Wang Jie. Screening of single-chain variable fragment (scFv) to bone sialoprotein[J]. Chinese journal of cellular and molecular immunology, 2011, 27(9): 979-981 |
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| Authors: | Wang Jiang-Tao Wang Jie |
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| Affiliation: | WANG Jiang-tao,WANG JieDepartment of Medical Research,Guangzhou General Hospital of Guangzhou Military Command,Guangzhou 510010,China |
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| Abstract: | AIM: To select single-chain variable fragment(scFv) antibody specific for bone sialoprotein(BSP) from Human Single Fold scFv Libraries.METHODS: Human Single Fold scFv Libraries were panned against immobilized BSP in a microtiter plate,after three rounds of panning,96clones were determined specific to BSP.The specificity of each scFv clone was determined by ELISA.The coding gene for BSP protein scFv has been sequenced.RESULTS: Phage antibody for BSP protein had a specific combination character.There were 368... |
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| Keywords: | bone sialoprotein(BSP) single-chain variable fragment(scFv) phage library |
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