Real-Time, label-free monitorine, of tumor antigen and serum antibody interactions
Real-Time, label-free monitorine, of tumor antigen and serum antibody interactions
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DOI：
10.1016/j.jbbm.2004.05.006
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发表时间：
2004-11-30
影响因子：
--
通讯作者：
Batt, CA
中科院分区：
文献类型：
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作者：
Campagnolo, C;Meyers, KJ;Batt, CA
检测生物分子相互作用的传统技术可能受到外源标记的需要的限制。时间和劳动密集的协议。也因为敏感程度差。本文设计了一种折射仪，通过表面等离子体共振（SPR）的变化来检测生物分子间的相互作用。根据表面异质性模型测定抗NY-ESO-1单克隆抗体ES121与癌睾丸抗原NY-ESO-1的结合动力学和亲和值，得到K-D值分别为1.3 x 10（-9）和2.1 x 10(-10) m。然后使用重新配置的仪器在预选的癌症患者血清样本中测量肿瘤抗原与针对这些抗原的血清抗体之间的相互作用。检测肿瘤抗原包括NY-ESO-1。SSX2和p53。所有的都用作含有多组氨酸标签的重组蛋白。这些结果表明，该仪器能够检测癌症患者血清前抗体与固定肿瘤抗原的结合，与先前基于elisa的实验结果一致。这些结果显示了SPR技术在快速诊断和监测免疫反应方面的潜力。(C) 2004 Elsevier B.V.版权所有
Conventional techniques for the detection of biomolecular interactions can be limited by the need for exogenous labels. time- and labor-intensive protocols. as well as by poor sensitivity levels. A refractometer instrument has been reconfigured to detect biomolecular interactions through changes ill surface plasmon resonance (SPR). The binding kinetics and affinity valuess of anti-NY-ESO-1 monoclonal antibody,, ES121, to the cancer-testis antigen NY-ESO-1 were determined according to the surface heterogeneity model and resulted in K-D values of 1.3 x 10(-9) and 2.1 X 10(-10) M. The reconfigured instrument was then used to measure the interaction between tumor antigens and serum antibodies against these antigens in preselected cancer patient sera samples. The tumor antigens assayed included NY-ESO-1. SSX2 and p53. all used as recombinant proteins containing polyhistidine tags. These results demonstrated that the instrument is capable of detecting the binding of serum antibodies front cancer patient sera to immobilized tumor antigens, consistent with those observed previously in ELISA-based experiments. These results demonstrate the potential of SPR technology for the rapid diagnosis and monitoring immune responses. (C) 2004 Elsevier B.V. All rights reserved.