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一种用于食品筛查的生物传感器微阵列的开发,采用成像表面等离子体共振技术。

Development of a biosensor microarray towards food screening, using imaging surface plasmon resonance.

作者信息

Rebe Raz Sabina, Bremer Maria G E G, Giesbers Marcel, Norde Willem

机构信息

RIKILT - Institute of Food Safety, Wageningen UR, P.O. Box 230, 6700 AE Wageningen, The Netherlands.

出版信息

Biosens Bioelectron. 2008 Dec 1;24(4):552-7. doi: 10.1016/j.bios.2008.05.010. Epub 2008 Jun 3.

Abstract

In this study we examined the possibilities of implementing direct and competitive immunoassay formats for small and large molecule detection on a microarray, using IBIS imaging surface plasmon resonance (iSPR) system. First, IBIS iSPR optics performance was evaluated. Using a glycerol calibration curve on underivatized surface we observed high baseline variability, but uniform and robust sensitivity between hundred regions of interest. Further on, a direct immunoassay for bovine IgG detection and a competitive immunoassay for gentamicin and neomycin were developed. The direct immunoassay for bovine IgG detection in a microarray format showed poor sensitivity in comparison to the assay performed in Biacore 3000, due to low immobilization efficiency on spots. The competitive immunoassay for parallel gentamicin and neomycin detection in a microarray format displayed sensitivity in the ngmL(-1) range, comparable with the sensitivity achieved in Biacore 3000 and in the range of maximum residue limits in milk, established in the European Union. We expect that, utilization of the IBIS iSPR system for food analysis, by screening high and low molecular weight compounds, will allow rapid and simultaneous detection of various ingredients and contaminants, providing the end-user with a detailed food profile. However, assay transfer from conventional SPR biosensors to the imaging microarray platform also presents new challenges, such as sufficient immobilization on spots, that must be addressed in future studies.

摘要

在本研究中,我们使用IBIS成像表面等离子体共振(iSPR)系统,研究了在微阵列上实现用于小分子和大分子检测的直接和竞争性免疫分析形式的可能性。首先,评估了IBIS iSPR光学性能。在未衍生化表面上使用甘油校准曲线,我们观察到基线变异性较高,但在一百个感兴趣区域之间具有均匀且稳健的灵敏度。进一步地,开发了用于检测牛IgG的直接免疫分析以及用于庆大霉素和新霉素的竞争性免疫分析。与在Biacore 3000中进行的分析相比,微阵列形式的用于检测牛IgG的直接免疫分析灵敏度较差,这是由于斑点上的固定效率较低。微阵列形式的用于同时检测庆大霉素和新霉素的竞争性免疫分析在ngmL(-1)范围内显示出灵敏度,与在Biacore 3000中实现的灵敏度相当,且在欧盟规定的牛奶最大残留限量范围内。我们期望,通过筛选高分子量和低分子量化合物,将IBIS iSPR系统用于食品分析,能够快速同时检测各种成分和污染物,为最终用户提供详细的食品概况。然而,从传统SPR生物传感器向成像微阵列平台的分析转移也带来了新的挑战,例如在斑点上的充分固定,这必须在未来的研究中加以解决。

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