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应用噬菌体随机肽库技术筛选SARS-CoVS蛋白模拟表位 被引量：1

Screening and identification of mimotope of the SARS-CoV spike protein by phage peptide library
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摘要 目的筛选SARS病毒(Severe acutere spiratorysyndromeas sociated coronavirus,SARSCoV)结构蛋白S(Spike)特异性的模拟表位,为抗SARS疫苗研究提供基础。方法以抗SARS病毒S蛋白的单克隆抗体为固相筛选分子,对人工合成的噬菌体随机12肽库进行5轮“吸附洗脱扩增”的筛选,随机挑选30个克隆,经噬菌体酶联免疫吸附(ELISA)法和交叉反应实验鉴定阳性克隆,再进行DNA序列分析和竞争抑制结合实验,以确定SARS病毒S蛋白的模拟表位。结果经噬菌体富集后,从随机挑选的30个克隆中得到了29个编码8种12肽的阳性克隆,8条肽与S蛋白的320～350氨基酸序列有较高同源性,确定YF、W、E和K氨基酸残基为模拟表位的骨架结构。结论用噬菌体12肽库成功筛选到了SARS病毒S蛋白的模拟表位,为基于S蛋白的肽疫苗研制提供了基础。 Objective To screen the mimotopes of SARS-CoV spike protein by using monoclonal antibody and phage peptide library. Methods By using SARS-CoV spike protein specific monoclonal antibody as selective molecule, a 12-mer phage peptide library was biopanned and positive clones were selected by ELISA,competition assay, and DNA sequencing. Results There were approximately 20000 times of enrichment about the titer of bound phages after five rounds biopanning procedures. Twenty-nine positive clones encoding 8 different 12-mer peptides were selected. These peptides were highly homologous with 320 - 350 amino acid sequence of SARS-CoV S protein, and most of the phage clones had the motifs of F/Y（X）WE（X）K. Conclusion SARS-CoV S protein mimotope is obtained by phage peptide library screening, which provides bases for anti-SARS-CoV peptide vaccine development.
作者 张小萍 王靖雪 魏静 万瑛 吴玉章
机构地区 第三军医大学全军免疫学研究所
出处 《免疫学杂志》 CAS CSCD 北大核心 2005年第6期442-444,452,共4页 Immunological Journal
基金 国家自然科学基金重大项目(30490240) 国家杰出青年科学基金(30325020)资助
关键词 SARS病毒 S蛋白 模拟表位 噬菌体肽库 SARS-CoV Spike protein Momitope Phage peptide library
分类号 R392.1 [医药卫生—免疫学]
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参考文献11

  • 1Drosten C,Gunther S,Preiser W,et al.Identification of a novel coronavirus in patients with severe acute respiratory syndrome [J].N Engl J Med,2003,348 (20):1967-1976.
  • 2Rota PA,Oberste MS,Monroe SS,et al.Characterization of a novel coronavirus associated with severe acute respiratory syndrome [J].Science,2003,300 (5 624):1394-1399.
  • 3Marra MA ,Jones SJ ,Astell CR ,et al .The genome sequence of the SARS-associated coronavirus [J].Science,2003,300 (5 624):1399-1404.
  • 4He Y,Zhou Y,Liu S,et al.Receptor-binding domain of SARS-CoV spike protein induces highly potent neutralizing antibodies:implication for developing subunit vaccine [J].Biochem Biophys Res Commun,2004,324 (2):773-781.
  • 5Scott JK,Smith GP.Searching for peptide ligands with an epitope library [J].Science,1990,249 (4967):386-390.
  • 6Wrighton NC,Farrel FX ,Chang R,et al.Small peptides as potent mimetics of the protein hormone erythropoietin [J].Science,1996,273 (5274):458-464.
  • 7Chang YJ,Liu CY,Chiang BL,et al.Induction of IL-8 release in lung cells via activator protein-1 by recombinant baculovirus displaying severe acute respiratory syndrome-coronavirus spike proteins:identification of two functional regions[J].JImmunol,2004,173 (12):7602-7614.
  • 8D'Mello F,Partidos CD,Steward MW,et al.Definition of the primary structure of hepatitis B virus (HBV) pre-S hepatocyte binding domain using random peptide libraries [J].Virology,1997,237 (2):319-326.
  • 9Scala G,Chen X,Liu W,et al.Selection of HIV-specific immunogenic epitopes by screening random peptide libraries with HIV-1-positive sera [J].J Immunol,1999,162 (10):6155-6161.
  • 10Li W,Moore MJ,Vasilieva N,et al.Angiotensin-converting enzyme 2 is a functional receptor for the SARS coronavirus [J].Nature,2003,426 (6 965):450-454.

同被引文献12

  • 1王国华,张贺秋,于继云,陈坤,宋晓国,许红莲,阎瑾琦,修冰水,何竟,邱艳,凌世淦.SARS冠状病毒S蛋白表位合成肽抗原性的研究[J].中华微生物学和免疫学杂志,2004,24(7):556-558. 被引量：3
  • 2张嘉杰,吴少瑜,徐伟,吴曙光.SARS-CoVS蛋白抗原表位多肽的设计合成与免疫反应[J].解放军药学学报,2005,21(1):40-42. 被引量：3
  • 3Peiris JSM, Yuen KY, Osterhaus ADME, et al. The severe acute respiratory syndrome [J]. N Engl J Med, 2003,349 ( 25 ) : 2431-2441.
  • 4Heike H, Kim H, Andrea M, et al. S protein of severe acute respiratory syndrome-a.ssociated eoronavirus mediates entry into hepatoma cell lines and is targeted by neutralizing antibodies in infected patients [J]. ] Virol, 2004,78(12):6134-6142.
  • 5Nie Y, Wang G, Shi X, et al. Neutralizing antibodies in patients with severe acute respiratory syndrome-associated coronavirus infection [J]. J Infect Dis, 2004, 190(6) : 1119- 1126.
  • 6Wong VW, Dai D, Wu AK, et al. Treatment of severe acute respiratory syndrome with convalescent plasma [J]. Hong Kong Med J, 2003,9(3) : 199-201
  • 7Yang ZY, Kong WP, Huang Y, et al. A DNA vaccine induces SARS coronavirus neutralization and protective immunity in mice [J]. Nature, 2004,428(6982) :561-564.
  • 8Kanta S, Josephine M, Leatrice V, et al. Prior infection and passive transfer of neutralization antibody prevent replication of severe acute respiratory syndrome eoronovirus in the respiratory tract of mice [J]. Virol, 2004,78(7):3572-3577.
  • 9Saha S, Bhasin M, Raghava GP. Bcipep: a database of B- cell epitopes [ J ]. BMC Genomics, 2005,6 ( 1 ) : 79.
  • 10Zhang H, Wang G, Li J, et al. Identification of an antigenic determinant on the S2 domain of the severe acute respiratory syndrome coronavirus spike glycoprotein capable of inducing neutralizing antibodies [J]. J Virol, 2004,78 (13) :6938-6945.

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免疫学杂志
2005年 第6期

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