Surface Plasmon Resonance Immunoassay for Biotin Determination on a Home-made Instrument

Procedia Technology Pub Date : 2017-01-01 DOI:10.1016/j.protcy.2017.04.037

Xing Chen, Lulu Zhang, Dafu Cui

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引用次数: 4

Abstract

In this paper, a home-made SPR analytical system and a denaturalized bovine serum albumin (dBSA) SPR chip were used to detect biotin based on the method of an inhibition immunoassay. The experimental was compared with a commercial protocol. The conditions of regeneration were optimized. The dBSA SPR chip was fabricated by a self-assembling method and the derivative of biotin was immobilized on the dBSA SPR chip by using a on-line mode. A commercial instrument and a dextran-SPR-chip (CM5) were used to implement the same experiments.

Based on the experiments of immune reactions between biotin antibody and biotin derivative, the biotin antibody concentration was optimized at about 5 μg/mL in order to obtain highly sensitivity, which was used in the followed inhibition immunoassays. The lowest detection limit for biotin is 0.1 μg/mL with a detection range of 0.1-1000 μg/ml by using our home-made instrument and the dBSA-SPR-chip. The detection results of our instrument and dBSA SPR chip are comparable with the commercial protocol. Our home-made instrument and dBSA-SPR-chips can be also applied to detect other small molecules based on the indirect inhibition immunoassay.

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表面等离子体共振免疫分析法在国产仪器上测定生物素

本文采用自制的SPR分析系统和变性牛血清白蛋白(dBSA) SPR芯片，采用抑制免疫分析法对生物素进行检测。实验与商业方案进行了比较。对再生条件进行了优化。采用自组装方法制备了dBSA SPR芯片，并通过在线方式将生物素衍生物固定在dBSA SPR芯片上。使用商用仪器和右旋糖酐- spr芯片(CM5)进行相同的实验。通过生物素抗体与生物素衍生物的免疫反应实验，优化生物素抗体浓度为5 μg/mL左右，获得高灵敏度，用于后续的抑制免疫试验。生物素的最低检出限为0.1 μg/mL，检测范围为0.1 ~ 1000 μg/mL。本仪器和dBSA SPR芯片的检测结果与商用协议相当。我们的自制仪器和dbsa - spr芯片也可以应用于间接抑制免疫分析法检测其他小分子。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Procedia Technology

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