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通过表面等离子体共振免疫分析法测定人血清样本中的人生长激素。

Determination of human growth hormone in human serum samples by surface plasmon resonance immunoassay.

作者信息

Treviño J, Calle A, Rodríguez-Frade J M, Mellado M, Lechuga L M

机构信息

Grupo de Nanobiosensores y Biofísica Molecular, Centro de Investigación en Nanociencia y Nanotecnología (CIN2: CSIC-ICN), ETSE, Campus UAB, Bellaterra, Barcelona, Spain.

出版信息

Talanta. 2009 May 15;78(3):1011-6. doi: 10.1016/j.talanta.2009.01.012. Epub 2009 Jan 20.

Abstract

A surface plasmon resonance immunoassay has been developed to determine human growth hormone (hGH) directly and without pre-treatment in human serum samples. A binding inhibition immunoassay was employed. Antibody concentration, assay buffer and regeneration solution have been optimized in order to reach the best performance and the lower non-specific binding of the matrix components to the sensor surface. The lowest detection limit was 6 ng/mL, with a working range covering the physiological range. Reproducibility of the assay was excellent with both intra-assay and inter-assay relative standard deviations <5%, while a variation of 2.19% was obtained employing different sensor chips. Reutilization of the sensor surface allows its continuous use over 50 measurements with a signal drop <20%. The SPR immunoassay results were validated using enzyme-linked immunosorbent assay (ELISA) showing an excellent correlation (R(2)=0.985). A portable and fully automated system (Sensia SL) was employed in this work. This is the first SPR biosensor assay capable of detecting relevant concentrations of a clinical analyte in serum. This study shows the potentials of this device as a diagnostic tool for the detection of multiple clinical analytes.

摘要

已开发出一种表面等离子体共振免疫分析法,可直接测定人血清样本中的人生长激素(hGH),且无需预处理。采用了结合抑制免疫分析法。对抗体浓度、分析缓冲液和再生溶液进行了优化,以实现最佳性能,并降低基质成分与传感器表面的非特异性结合。最低检测限为6 ng/mL,工作范围涵盖生理范围。该分析方法的重现性极佳,批内和批间相对标准偏差均<5%,而使用不同的传感器芯片时变异系数为2.19%。传感器表面的可重复使用性使其能够连续进行50多次测量,信号下降<20%。使用酶联免疫吸附测定法(ELISA)对表面等离子体共振免疫分析结果进行了验证,显示出极佳的相关性(R(2)=0.985)。本研究中使用了一种便携式全自动系统(Sensia SL)。这是首个能够检测血清中临床分析物相关浓度的表面等离子体共振生物传感器分析方法。本研究展示了该设备作为检测多种临床分析物的诊断工具的潜力。

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