Baek Taek Jin, Kim Nam Hyun, Choo Jaebum, Lee Eun Kyu, Seong Gi Hun
Department of Applied Chemistry, Hanyang University, Ansan 425-791, Korea.
J Microbiol Biotechnol. 2007 Nov;17(11):1826-32.
We describe the fabrication of poly(ethylene glycol) diacrylate (PEG-DA) hydrogel microstructures with a high aspect ratio and the use of hydrogel microstructures containing the enzyme beta-galactosidase (beta-Gal) or glucose oxidase (GOx)/horseradish peroxidase (HRP) as biosensing components for the simultaneous detection of multiple analytes. The diameters of the hydrogel microstructures were almost the same at the top and at the bottom, indicating that no differential curing occurred through the thickness of the hydrogel microstructure. Using the hydrogel microstructures as microreactors, beta-Gal or GOx/HRP was trapped in the hydrogel array, and the time-dependent fluorescence intensities of the hydrogel array were investigated to determine the dynamic uptake of substrates into the PEG-DA hydrogel. The time required to reach steady-state fluorescence by glucose diffusing into the hydrogel and its enzymatic reactions with GOx and HRP was half the time required for resorufin beta-D-galactopyranoside (RGB) when used as the substrate for beta-Gal. Spatially addressed hydrogel microarrays containing different enzymes were micropatterned for the simultaneous detection of multiple analytes, and glucose and RGB solutions were incubated as substrates. These results indicate that there was no cross-talk between the beta-Gal-immobilizing hydrogel micropatches and the GOx/HRP-immobilizing micropatches.
我们描述了具有高纵横比的聚乙二醇二丙烯酸酯（PEG-DA）水凝胶微结构的制备，以及使用含有β-半乳糖苷酶（β-Gal）或葡萄糖氧化酶（GOx）/辣根过氧化物酶（HRP）的水凝胶微结构作为生物传感组件来同时检测多种分析物。水凝胶微结构的顶部和底部直径几乎相同，这表明在水凝胶微结构的厚度方向上没有发生差异固化。将水凝胶微结构用作微反应器，将β-Gal或GOx/HRP捕获在水凝胶阵列中，并研究水凝胶阵列随时间变化的荧光强度，以确定底物向PEG-DA水凝胶中的动态摄取。葡萄糖扩散到水凝胶中并与GOx和HRP发生酶促反应达到稳态荧光所需的时间，是间苯二酚β-D-吡喃半乳糖苷（RGB）用作β-Gal底物时所需时间的一半。对包含不同酶的空间寻址水凝胶微阵列进行微图案化，以同时检测多种分析物，并将葡萄糖和RGB溶液作为底物进行孵育。这些结果表明，固定有β-Gal的水凝胶微斑与固定有GOx/HRP的微斑之间没有串扰。
