疏水蛋白
微流控
涂层
基质(水族馆)
单层
色谱法
材料科学
化学
接触角
纳米技术
分析化学(期刊)
生物化学
复合材料
基因
海洋学
地质学
作者
Jan-Niklas Klatt,Tobias Hutzenlaub,Thomas Subkowski,Tanja Müller,Stefan Hennig,Roland Zengerle,Nils Paust
出处
期刊:ACS applied polymer materials
[American Chemical Society]
日期:2021-06-07
卷期号:3 (7): 3278-3286
被引量:3
标识
DOI:10.1021/acsapm.0c01301
摘要
Microfluidics allows the miniaturization of biochemical analyses. Small dimensions reduce sample and reagent consumption and enhance reaction rates. A downside is that high surface-to-volume ratios increase the unspecific binding of proteins to the substrate material. The resulting sample loss and reagent depletion decrease the sensitivity and specificity of protein-based assays, especially if low concentrations are analyzed. Here, we introduce the hydrophobin coating of microfluidic chips made of cyclic olefin copolymers (COC). The recombinant hydrophobin H*Protein B self-assembles into stable monolayers on hydrophobic surfaces, making them hydrophilic and thus reducing hydrophobic interactions between the chip surfaces and proteins. The substrate and sealing layers of the microfluidic chip were simply dip-coated and subsequently assembled by thermodiffusion bonding, which renders our coating procedure compatible with mass fabrication. Contact angle measurements and atomic force microscopy were used to evaluate the effect of high temperatures (107 °C) on COC substrates coated with H*Protein B. The efficiency of the protein-repellent coating was evaluated by depletion experiments with bovine serum albumin, human serum, and cerebrospinal fluid in microfluidic chips. Protein recovery was investigated down to protein concentrations of 0.3 μg/mL. Recoveries of 90% were observed with total protein amounts of 10 ng, even for microfluidic channels up to 835 mm in length and with a cross section of 80 μm × 230 μm in a COC 6013/8007 foil. For comparison, only 30 and 60% of the protein was recovered in uncoated microfluidic channels with lengths of 835 and 128 mm, respectively. The long-term stability of the hydrophobin-coated chips for 8 weeks was demonstrated.
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