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利用噬菌体肽库确定IL-2Rα表位序列

Identification of IL-2Ra epitope sequences by phage peptide library

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【作者】 刘北一; 李一; 王莉; 于春雷; 杨贵贞; 富宁; 周长城; 齐杰; 周慧; 李惟;

【Author】 LIU Bei-Yi;LI Yi ;WANG Li et al (Department of Immunology, Norman Bethune University of Medical Sciences,Changchun 130021)

【机构】 白求恩医科大学免疫学教研室!长春130021; 第一军医大学免疫学教研室!广州510;

【摘要】 目的:确定IL-2Rα表位,为研制高效特异性强的小分子肽类免疫抑制剂奠定基础。方法:用抗IL-2Rα单克隆抗体5G1对噬菌体六肽库进行筛选,选择出与5G1结合较强的克隆,经DNA序列分析,获得保守序列。对合保守序列的克隆进行生物学活性鉴定。结果:选择出31个与5G1结合较强的克隆。获得Ser-Ser-Phe和Ser-Ser-Arg两种保守序列。生物学活性鉴定表明:含Ser-Ser-Arg序列克隆较含Ser-Ser-Phe被抑制效果好。结论:Ser-Ser-Arg是IL-2Rα表位的关键序列。以此表位序列合成的小分子肽可作为IL-2Rα的拮抗剂而成为免疫抑制剂。

【Abstract】 Objective: TO determine the amino acid sequences Of IL-2R epitope that would be useful in making small peptide drugs asImmunosuppressant. Methods: A mouse monoclonal antibody that is specific for the human IL-2Ra 5G1 (anti Tac antibody) was used to screena random phage peptide library which have 6 amino-acid residues displayed. Phage clones which could highly react with 5G1 was selected. After DNA sequencing, conservative sequences were acquired. The biological activities of Phage clones were studied by sIL-2Ra blocking assay.Results: After 4 round of biopanning 31 phage clones were selected; two kinds of conservative sequence(Ser-Ser-Phe and Ser-Ser-Arg) weredetermined by single-strand dideoxy-sequencing by chine termination method. Ser-Ser-Arg clones were more effective than Ser-Ser-Phe clones.Conclusion:Ser-Ser -Arg seqience is the IL-2R epitope.

【基金】 卫生部科学研究基金!No 98-1-203
  • 【文献出处】 中国免疫学杂志 ,CHINESE JOURNAL OF IMMUNOLOGY , 编辑部邮箱 ,1999年12期
  • 【被引频次】4
  • 【下载频次】40
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