Niotis Aristidis E, Mastichiadis Christos, Petrou Panagiota S, Christofidis Ion, Siafaka-Kapadai Athanasia, Misiakos Konstantinos, Kakabakos Sotirios E
Immunoassay-Immunosensors Laboratory, Institute of Radioisotopes & Radiodiagnostic Products, N.C.S.R. Demokritos, Aghia Paraskevi, 15310, Athens, Greece.
Anal Bioanal Chem. 2009 Feb;393(3):1081-6. doi: 10.1007/s00216-008-2501-2. Epub 2008 Nov 11.
An optical capillary waveguide fluoroimmunosensor based on glass capillaries internally coated with an ultrathin poly(dimethylsiloxane) (PDMS) film is presented. The evaluation of the capillaries developed was done in comparison with aminosilanized [3-(aminopropyl)triethoxysilane, APTES] glass and poly(methylpentene) (PMP) capillaries by immobilizing rabbit gamma-globulins on the internal capillary wall. Following reaction with (R)-phycoerythrin-labelled antibody, the capillary was scanned with a laser beam and the fluorescence waveguided through the capillary wall was detected by a photomultiplier placed at one of its ends. The capillaries developed provided considerably improved protein coating homogeneity (intracapillary coefficients of variation 2.9-6.6%) and repeatability (intercapillary coefficients of variation 2.1-5.0%) compared with APTES-treated ones (7.9-13.4 and 8.5-15.2%, respectively). With use of these capillaries in a sandwich-type immunosensor for the determination of rabbit gamma-globulins, the assay detection limit was improved eightfold (4.4 ng/mL) compared with that obtained using PMP capillaries (35.3 ng/mL), whereas the assay repeatability was improved threefold (intra-assay coefficients of variation 5.9-13.1%) compared with APTES-treated capillaries (15.6-36%).
本文介绍了一种基于玻璃毛细管的光学毛细管波导荧光免疫传感器，该毛细管内部涂有超薄聚二甲基硅氧烷（PDMS）膜。通过将兔γ球蛋白固定在内壁上，将所开发的毛细管与氨基硅烷化（3-氨丙基三乙氧基硅烷，APTES）玻璃毛细管和聚甲基戊烯（PMP）毛细管进行比较评估。与（R）-藻红蛋白标记的抗体反应后，用激光束扫描毛细管，并通过放置在其一端的光电倍增管检测通过毛细管壁传导的荧光。与APTES处理的毛细管（分别为7.9-13.4%和8.5-15.2%）相比，所开发的毛细管提供了显著改善的蛋白质包被均匀性（管内变异系数为2.9-6.6%）和重复性（管间变异系数为2.1-5.0%）。在用于测定兔γ球蛋白的夹心型免疫传感器中使用这些毛细管时，与使用PMP毛细管（35.3 ng/mL）相比，检测限提高了八倍（4.4 ng/mL），而与APTES处理的毛细管（15.6-36%）相比，测定重复性提高了三倍（批内变异系数为5.9-13.1%）。
