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Construction and screening of human vitiligo phage autoantibody library based on in-cell PCR
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 16
- DOI:
- 10.3321/j.issn:1000-2790.2006.16.002
- Key Word:
- 白癜风;聚合酶链反应;噬菌体抗体库;自身抗体
Abstract： 目的: 克隆白癜风患者淋巴细胞中原始配对的自身抗体基因,构建噬菌体抗体库并筛选针对黑素细胞膜抗原的特异性抗体. 方法: 细胞内PCR法扩增人免疫球蛋白轻链和重链可变区基因,Cre重组酶介导下完成单个细胞内的轻重链连接,获得反映体内原始配对信息的单链抗体可变区基因,构建噬菌体抗体库,筛选针对黑素细胞膜抗原的自身抗体. 结果: 构建完成库容量约为3×l06的噬菌体抗体库,筛选得到1株针对黑素细胞膜抗原的自身抗体. 结论: 利用细胞内PCR技术和噬菌体抗体库技术成功构建免疫文库,为进一步研究自身抗体在白癜风发生发展过程中的作用奠定了基础.
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