Derveaux Stefaan, Stubbe Barbara G, Roelant Chris, Leblans Marc, De Geest Bruno G, Demeester Jo, De Smedt Stefaan C
Laboratory of General Biochemistry and Physical Pharmacy, University of Ghent, Harelbekestraat 72, 9000 Ghent, Belgium.
Anal Chem. 2008 Jan 1;80(1):85-94. doi: 10.1021/ac071212i. Epub 2007 Dec 4.
The "layer-by-layer" (LbL) technology has been widely investigated for the coating of flat substrates and capsules with polyelectrolytes. In this study, LbL polyelectrolyte coatings applied at the surface of digitally encoded microcarriers were evaluated for the quantitative, sensitive, and simultaneous detection of proteins in complex biological samples like serum, plasma, and blood. LbL coated microcarriers were therefore coupled to capture antibodies, which were used as capture agents for the detection of tumor necrosis factor (TNF-alpha), P24, and follicle stimulating hormone (FSH). It was found that the LbL coatings did not disassemble upon incubating the microcarriers in serum and plasma. Also, nonspecific binding of target analytes to the LbL coating was not observed. We showed that the LbL coated microcarriers can reproducibly detect TNF-alpha, P24, and FSH down to the picogram per milliliter level, not only in buffer but also in serum and plasma samples. Microcarrier-to-microcarrier intratube variations were less then 30%, and interassay variations less than 8% were observed. This paper also shows evidence that the LbL coated digitally encoded microcarriers are ideally suited for assaying proteins in "whole" blood in microfluidic chips, which are of high interest for "point-of-care" diagnostics.
“逐层”(LbL)技术已被广泛研究用于用聚电解质涂覆平板基材和胶囊。在本研究中,评估了应用于数字编码微载体表面的LbL聚电解质涂层对血清、血浆和血液等复杂生物样品中蛋白质的定量、灵敏和同时检测能力。因此,LbL包被的微载体与捕获抗体偶联,这些捕获抗体用作检测肿瘤坏死因子(TNF-α)、P24和促卵泡激素(FSH)的捕获剂。研究发现,将微载体在血清和血浆中孵育时,LbL涂层不会解体。此外,未观察到目标分析物与LbL涂层的非特异性结合。我们表明,LbL包被的微载体不仅可以在缓冲液中,而且可以在血清和血浆样品中,以皮克每毫升的水平可重复地检测TNF-α、P24和FSH。观察到微载体间管内差异小于30%,批间差异小于8%。本文还表明,LbL包被的数字编码微载体非常适合在微流控芯片中对“全”血中的蛋白质进行检测,这对于“即时护理”诊断具有重要意义。