表面等离子共振法测?茁-淀粉样多肽人源性单链抗体E3 scfv的亲和力
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江苏省教育厅产业化项目(JH01-048)


Determination of affinity of human single-chain variable fragment antibody against ?茁-amyloid peptide by SPR
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    摘要:

    目的:采用表面等离子共振(surface plasmon resonance,SPR)生物传感器测定-茁-淀粉样多肽(A-茁)人源性单链抗体E3 scfv的亲和力-方法:将A-茁1-40固定于传感芯片表面,以E3 scfv等抗体为流动相,实时监测抗原抗体的结合和解离过程,并对其相互作用模式和动力学常数进行分析-结果:E3 scfv-阳性对照抗体DE2B4均能与A-茁1-40发生特异性免疫结合反应,阴性对照抗体9E10则不能结合A-茁1-40;A-茁1-40与E3 scfv及DE2B4的结合呈现快结合慢解离的动力学过程,其中DE2B4与A-茁1-40的亲和力为6.77 × 10-7 mol/L,E3 scfv的亲和力为5.38 × 10-6mol/L-结论:SPR适宜用作测定E3 scfv与A-茁1-40的亲和力-

    Abstract:

    Objective:To investigate the affinity of human single-chain variable fragment antibody against -茁-amyloid peptide(A-茁) by surface plasmon resonance(SPR) sensor. Methods:A-茁1-40 was fixed on sensing chip surface and the antibodies as E3 scfv were applied as mobile phase. Binding and separation of A-茁1-40 with the antibodies were monitored in real time. The interaction mode between antigen and antibody and the relevant dynamic parameters were determined. Results:A specific interaction between A-茁1-40 and E3 scfv or DE2B4,rather than 9E10 was detected. The binding and separation between the antigen and antibody were dynamic with a slow speed; Affinity DE2B4 with A-茁1-40 was 6.77 × 10-7 mol/L and that of E3 scfv was 5.38 × 10-6 mol/L. Conclusion:SPR is useful in determination of affinity of E3 scfv with A-茁1-40.

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解 鹏,李 玥,陈 琪.表面等离子共振法测?茁-淀粉样多肽人源性单链抗体E3 scfv的亲和力[J].南京医科大学学报(自然科学版),2008,28(11):1389-1392

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  • 收稿日期:2008-07-20
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