期刊文献+

抗人凝血因子Ⅷ-C1C2区单克隆抗体的制备及鉴定 被引量:1

Preparation and Characterization of Monoclonal Antibody to FⅧ C1C2 Domain of Human
原文传递
导出
摘要 目的制备抗人凝血因子Ⅷ(FⅧ)C1C2区单克隆抗体(mAb),并对其进行生化和生物学鉴定。方法采用基因重组技术表达FⅧ-C1C2区蛋白(rFⅧ-C1C2),以rFⅧ-C1C2免疫8周龄Balb/C小鼠,小鼠脾脏细胞与小鼠骨髓瘤细胞融合后,ELISA法筛选阳性克隆,制备稳定分泌抗rFⅧ-C1C2mAb的细胞株,并对mAb进行生化和免疫学鉴定。结果获得一株抗rFⅧ-C1C2区单克隆抗体阳性克隆2B11,命名为SZ-133。琼脂糖双扩散鉴定为IgG1类,Westernblot法证实SZ-133可与rFⅧ-C1C2区蛋白及重组的全长FⅧ特异性结合,但不影响FⅧ的凝血活性。结论表达了rFⅧ-C1C2,并制备出了抗人凝血因子Ⅷ的单克隆抗体SZ-133,它能特异识别血浆及重组的FⅧ,有可能用于基础和临床应用研究。 Objective To develop monoclonal antibody(mAb)anti-FⅧ C1C2 domain of human and characterize its biochemical and biological functions.Methods FactorⅧ-C1C2 protein was expressed by prokaryotic expression system.Balb/c mice of eight-week ages were immunized by intraperitoneal and subcutaneous injection of rFⅧ-C1C2 protein.The positive clone was screened by ELISA and identified using western-blot.Results A positive clone 2B11 designated as SZ-133 was produced,belonging to IgG1 subclass.Western blot demonstrated that SZ-133 could specifically recognized rFⅧ-C1C2 and fⅧ in human plasma.Conclusion rFⅧ-C1C2 is expressed in Ecoli.The successfully developed mAb against human FⅧ-C1C2 domain mAb can be useful for basic and clinical application.
出处 《苏州大学学报(医学版)》 CAS 北大核心 2010年第1期106-109,共4页 Suzhou University Journal of Medical Science
基金 江苏省高校自然科学基金资助项目(08KJD310008) 江苏省卫生厅"科教兴卫工程"临床医学中心血液病开放课题基金资助项目(WKF07009)
关键词 人凝血因子Ⅷ(FⅧ)C1C2区 原核表达 单克隆抗体 鉴定 human coagulation factor Ⅷ(FⅧ)C1C2 domain prokaryotic expression monoclonal antibody identification
  • 相关文献

参考文献8

  • 1Vehar GA, Keyl B, Eaton D, et al. Structure of human factor Ⅷ. Nature. 1984,312(22) :337 -342.
  • 2Henriet Meemsl, Alexander B Meijerl, Dave Cullinan,et al. Factor ⅧC1 domain residues 2092 -2093 participate in membrane binding[ J ]. Blood ,2008,114 ( 18 ) :587.
  • 3Ting-Chang Hsu, Kathleen P Pratt,Arthur R. Thompson, et al. FⅧ C1 domain loop containing phe2093 contributes affinity to the binding of recombinant FⅧC1 C2 proteins to activated platelets. Blood, 2008,112 ( 1 ) : 3069.
  • 4Ting-Chang Hsu, Kathleen P Pratt, Arthur R Thompson, et al. The factor Ⅷ C1 domain contributes to platelet binding [J]. Blood,2008,111 ( 1 ) :200 -208.
  • 5范磊,沈飞,谢丽倩,周梦云,阮长耿(指导).Mer免疫球蛋白样区原核蛋白表达和单克隆抗体制备[J].苏州大学学报(医学版),2007,27(1):4-7. 被引量:3
  • 6阮长耿 奚晓东 李佩霞 等.抗人血小板单克隆抗体的研究,一组分泌抗人血小板单克隆抗体的小鼠杂交瘤细胞.中华血液学杂志,1985,6(2):130-133.
  • 7Marc Jacquemin, Abdellah Benhida, Kathelijne Peerlinck, et al. Ahuman antibody directed to the factor Ⅷ Cl domain inhibits factor Ⅷ cofactor activity and binding to von- Willebrand factor, blood. 2000,95( 1 ) :156 - 163.
  • 8Steven W Pipe, Randal J Kaufman. Factor Ⅷ C2 domain missense mutations exhibit defective trafficking of biologically functional proteins. The Journal of Biological Chemistry,1996,271 (41) :25671 -25676.

二级参考文献9

  • 1Manfioletti G,Brancolini C,Avanzi G,et al.The protein encoded by a growth arrest-specific gene (gas 6) is a new member of the vitamin K-dependent proteins related to protein S,a negative coregulator in the blood coagulation cascade[J].MCB,1993,13(8):4976-4985.
  • 2Douglas K Graham,Thomas L Dawson,David L Mullaney,et al.Clonin8 and mRNA expression analysis of a novel human protooncogene,c-uer.Cell Growth & Differentiation,1994,5 (9):647-657.
  • 3阮长耿 奚晓东 李佩霞 等.抗人血小板单克隆抗体的研究,一组分泌抗人血小板单克隆抗体的小鼠杂交瘤细胞.中华血液学杂志,1985,6(2):130-133.
  • 4Toru Nakano,Junji Kisshino,Hitoshi Arita.Characterization of a high-affinity and specific binding site for Gas 6[J].FEBS,1996,387(1):75 -77.
  • 5Katherine L Guttridge,J Christopher Luft,Thomas L Dawson,et al.Mer receptor tyrosine kinase signaling prevention of apoptosis and alteration of cytoskeletal architecture without stimulation or proliferation[J].JBC,2002,277 (27):24057-24066.
  • 6Todd D Camenisch,Beverly H Koller,H Shelton Earp,et al.A novel receptor tyrosine kinase,Mer,inhibits TNF-α production and lipopolysaccharide-induced endotoxic shock.The Journal of Immunology,1999,162(6):3498-3503.
  • 7Anne Angelillo-Scherrer,Pablo Garcia de Frutos,Cristina Aparicio,et al.Deficiency or inhibition of Gas 6 causes platelet dysfunction and protects mice against thrombosis[J].Nature Medicine,2001,7 (2):215-221.
  • 8Margherita Gallicchio,Stefania Mitola,Donatella Valdembri,et al.Inhibition of vascular endothelial growth factor receptor 2-mediated endothelial cell activation by Axl tyrosine kinase receptor[J].Blood.2005,105(5):1970-1976.
  • 9Sara EK,Hogerkorp CM,Dictor M,et al.Mantle cell lymphomas express a distinct genetic signature affecting lymphocyte trafficking and growth regulation as compared with subpopulations of normal human B cells[J].Cancer Res,2002,62 (15):4398-4405.

共引文献5

同被引文献14

  • 1阮长耿,吴德沛,王兆钺,等.现代血液病诊断治疗学,第1版.安徽:科学技术出版社,2006:239-242.
  • 2Sahud MA,Pratt KP,Zhukov O,et al.ELISA system for detection of immune responses to factor Ⅷ:a study of 246 samples and correlation with the Bethesda assay.Haemophilia,2007; 13(3):317-322.
  • 3Blanco AN,Peirano AA,Grosso SH,et al.An ELISA system to detection anti-factor Ⅷ antibodies without interference by lupus anticoagulants.Haematologica,2000; 85(10):1045-1050.
  • 4Lindgem A,Wadenvik H,Tengbom L.Characterization of inhibitors to factor Ⅷ with an ELISA in congenital and acquired hemophilia A.Haemophilia,2002 ; 8 (5):644-648.
  • 5Jolanta KA,Behnaz PS,Saulius B,et al.Quantitation of antifactor Ⅷ antibodies in human plasma.Blood,2009; 113 (11):2587-2594.
  • 6Kopecky EM,Greinstetter S,Pabinger I,et al.Mapping of FⅧ inhibitor epitopes using cellulose-bound synthetic peptide arrays.J Immunol Methods,2006 ; 308 (1-2):90-100.
  • 7Scandella DH,Nakai H,Felch M,et al.In Hemophilia A and Autoantibody Inhibitor Patients:the Factor Ⅷ A2 Domain and Light Chain Are Most Immunogenic.Thromb Res,2001 ; 101 (5):377-387.
  • 8Christine L,Kempton C,Gilbert C,et al.How we treat a hemophilia A patient with a factor Ⅷ inhibitor.Blood,2009; 113(1):11-17.
  • 9Gharagozlous S,Sharifian RA,Khoshnoodi J,et al.Epitope specificity of anti-factor Ⅷ antibodies from inhibitor positive acquired and congenital haemophilia A patients using synthetic peptides spanning A and C domains.Thromb Haemost,2009 ; 101(5):834-839.
  • 10Pier MM.Treatment of von Willebrand's Disease.N Engl J Med,2004; 351(7):683-694.

引证文献1

二级引证文献2

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部