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检测可溶型CD226双mAb夹心ELISA的建立及初步应用 被引量:3

Development and preliminary application of a double mAb sandwich ELISA for detecting soluble CD226
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摘要 目的 :建立一种检测可溶型CD2 2 6 (sCD2 2 6 )的双mAb夹心ELISA ,并用于临床标本的检测。方法 :确定包被mAbLeoA1和HRP mAbFMU3的最佳工作浓度及最佳稀释液后 ,建立双mAb夹心ELISA ,并对方法的特异性、敏感性和稳定性进行鉴定。再以建立的方法检测临床标本。结果 :包被mAbLeoA1的最佳工作浓度为 2 .5mg/L ,HRP mAbFMU3的最佳工作浓度 1∶4 0 0 ,标准品及HRP mAb的最佳稀释液分别为 1g/LBSA 1mL/LTween2 0 PBS和 30 g/LPEG PBS .建立的双mAb夹心ELISA ,其特异性 (与人IgG无交叉反应 ,阻断实验中阻断效应呈剂量依赖性 )、敏感性 (检出下限为 110ng/L标准品CD2 2 6 /Fc融合蛋白 )及稳定性 (CV <10 .2 % )均较良好。对部分临床标本的检测中 ,发现 6例类风湿关节炎(RA)患者血清sCD2 2 6的水平升高 ,与正常人血清sCD2 2 6水平相比差异显著 (P <0 .0 1)。结论 :建立一种特异性高、重复性好、较敏感的检测sCD2 2 6的双mAb夹心ELISA 。 AIM: To develop a double mAb sandwich ELISA for detecting sCD226 and apply it to clinical specimens. METHODS: The optimal concentrations of coating mAb and HRP mAb, and the optimal diluents were determined. A double mAb sandwich ELISA then was established. The specificity, sentivity and stability of the method were identified. And clinical specimens were detected by this method. RESULTS: The optimal concentrations of coating mAb LeoA1 and HRP mAb FMU3 were 2.5 mg/L and 1∶400, respectively, and the optimal diluents for standard antigen and HRP mAb were 1 g/L BSA 1 mL/L Tween20 PBS and 30 g/L PEG PBS.The ELISA achieved satisfactory results in respect to specificity (no cross reaction to human IgG, in block test blocking effect was does dependent), sensitivity (the detected threshold was 110 ng/L) and stability (CV<10.2%). Statistical analysis showed that the serum sCD226 level in patients with rheumatoid arthritis (RA) was significantly higher than that in healthy adults. CONCLUSION: A double mAb sandwich ELISA with high specifity, and relative sensitivity for detection of sCD226 has been developed. It is shown to be an applicable method of detecting sCD226 in clinical specimens.
出处 《细胞与分子免疫学杂志》 CAS CSCD 北大核心 2003年第2期192-194,共3页 Chinese Journal of Cellular and Molecular Immunology
基金 国家重点基础研究发展规划资助项目 (No.2 0 0 1CB51 0 0 0 4 ) 国家自然科学基金重点资助项目 (No.30 0 30 1 30 )
关键词 可溶型CD226 单克隆抗体 酶联免疫吸附试验 临床应用 类风湿性关节炎 soluble sCD226(PTA1) ELISA rheumatoid arthritis
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  • 1Sherrington P D,J Bio Chem,1997年,272卷,21735页
  • 2Huang C,J Infect Dis,1994年,169卷,4期,868页
  • 3Scott J L,J Bio Chem,1989年,264卷,13475页

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  • 1Scott JL,Dunn SM,Jin BQ,et al.Characterization of a novel membrane glycoprotein involved in pl-atelet activation[J].Biol Chem,1989,264(23):13475-13482.
  • 2Sherrington PD,Scott JL,Jin BQ,et al.TLiSA1 (PTA1) activation antigen implicated in T cell differ-entiation and platelet activation is a member of the immunoglobulin superfamily exhibiting distinc-tive regulation of expression[J].Biol Chem,1997,272(35):21735-21744.
  • 3Tian F,Li D,Xia H,et al.Isolation of cDNAs encoding gibbon and monkey platelet and T cell activation antigen 1 (PTA1)[J].DNA Sequence,1999,10(3):155-161.
  • 4Wang PL,O' Farrell S,Clayberger C,et al.Identification and molecular cloning of tactile.A novel human T cell activation antigen that is a member of the Ig gene sperfamily[J].Immunol,1992,148(8):2600-2608.
  • 5Sloan KE,Eustace BK,Stewart JK,et al.CD155/PVR plays a key role in cell motility during tumor cell invasion and migration[J].BMC Cancer,2004,4(1):73-78.
  • 6Ralston KJ,Hird SL,Zhang XH,et al.The LFA-1-associated molecule PTA1 (CD226) on T cells forms a dynamic molecular complex with protein 4.1G and human discs large[J].Biol Chem,2004,279(32):33816-33828.
  • 7Shirakawa JSK,Shibuya A.Requiremengt of the serine at residue 329 for lipid raft recuitment of DNAM-1(CD226)[J].Int Immunol,2005,17(3):217-223.
  • 8Brycesom YT,March ME,Ljunggren HG,et al.Synergy among receptors of resting NK cells for the activation of natural cytotoxicity and cytokine secretion[J].Blood,2006,107(1):159-166.
  • 9Ma D,Sun Y,Lin D,et al.CD226 is expressed on the megakaryocytic lineage from hematopoietic stem cells/progenitor cells and involved in its polyploidization[J].Eur Haematol,2005,74:228-240.
  • 10EL-Sher4iny YM,Meade JL,Holmes TD,et al.The requirment for DNAM-1,NKG2D,anf NKp46 in the natural killer cell-mediated killing of myelima cells[J].Cancer Res,2007,67(18):8444-8449.

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