基于表面等离子体共振的降钙素原免疫测定法。

Surface plasmon resonance-based immunoassay for procalcitonin.

作者信息

Vashist Sandeep Kumar, Schneider E Marion, Barth Eberhard, Luong John H T

机构信息

Institute of Clinical Chemistry and Pathobiochemistry, Klinikum rechts der Isar der Technischen Universität München, Ismaninger Str. 22, D-81675, Munich, Germany.

Sektion Experimentelle Anaesthesiologie, University Hospital Ulm, Albert Einstein Allee 23, 89081 Ulm, Germany.

出版信息

Anal Chim Acta. 2016 Sep 28;938:129-36. doi: 10.1016/j.aca.2016.08.007. Epub 2016 Aug 9.

DOI:10.1016/j.aca.2016.08.007

PMID:27619095

Abstract

A surface plasmon resonance (SPR) biosensor has been developed for rapid immunoassay of procalcitonin (PCT) with high detection sensitivity and reproducibility. The 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC)-activated protein A (PrA), diluted in 1% (v/v) 3-aminopropyltriethoxysilane (APTES) was dispensed on a KOH-treated Au-coated SPR chip, resulting in the covalent binding of PrA in 30 min. This "single-step" PrA immobilization strategy led to the oriented binding of the anti-PCT antibody (Ab) on a PrA-functionalized gold (Au) chip. The leach-proof immobilization procedure is five-fold faster than conventional counterparts, enabling high detection specificity and reproducibility. The IA detects 4-324 ng mL(-1) of PCT with a limit of detection (LOD) and a limit of quantification (LOQ) of 4.2 ng mL(-1) and 9.2 ng mL(-1), respectively. It was capable of detecting PCT in real sample matrices and patient samples with high precision. The Ab-bound SPR chips were stable for more than five weeks.

摘要

已开发出一种表面等离子体共振（SPR）生物传感器，用于降钙素原（PCT）的快速免疫测定，具有高检测灵敏度和重现性。将用1%（v/v）3-氨丙基三乙氧基硅烷（APTES）稀释的1-乙基-3-（3-二甲基氨基丙基）-碳二亚胺盐酸盐（EDC）活化的蛋白A（PrA）滴加在经KOH处理的金涂层SPR芯片上，30分钟内实现PrA的共价结合。这种“一步法”PrA固定化策略使抗PCT抗体（Ab）在PrA功能化的金（Au）芯片上定向结合。这种防渗漏固定程序比传统方法快五倍，具有高检测特异性和重现性。该免疫测定法可检测4-324 ng mL⁻¹的PCT，检测限（LOD）和定量限（LOQ）分别为4.2 ng mL⁻¹和9.2 ng mL⁻¹。它能够高精度地检测实际样品基质和患者样品中的PCT。与抗体结合的SPR芯片稳定性超过五周。