Grup de Sensors i Biosensors, Departament de Química, Facultat de Ciències, Edifici C-Nord, Universitat Autònoma de Barcelona, 08193 Bellaterra (Cerdanyola del Vallès), Spain.
Anal Chem. 2010 Feb 1;82(3):1006-11. doi: 10.1021/ac902267f.
Low-temperature cofired ceramics (LTCC) technology is a versatile fabrication technique used to construct microflow systems. It permits the integration of several unitary operations (pretreatment, separation, (bio)chemical reaction, and detection stage) of an analytical process in a modular or monolithic way. Moreover, because of its compatibility with biological material, LTCC is adequate for analytical applications based on enzymatic reactions. Here we present the design, construction, and evaluation of a LTCC microfluidic system that integrates a microreactor (internal volume, 24.28 microL) with an immobilized beta-galactosidase from Escherichia coli (0.479 activity units) and an optical flow cell to measure the product of the enzymatic reaction. The enzyme was immobilized on a glyoxal-agarose support, maintaining its activity along the time of the study. As a proof of concept, the LTCC-beta-galactosidase system was tested by measuring the conversion of ortho-nitrophenyl beta-D-galactopyranoside, the substrate usually employed for activity determinations. Once packed in a monolithically integrated microcolumn, the miniaturized flow system was characterized, the operational conditions optimized (flow rate and injection volume), and its performance successfully evaluated by determining the beta-galactosidase substrate concentration at the millimolar level.
低温共烧陶瓷（LTCC）技术是一种通用的制造技术，用于构建微流系统。它允许以模块化或整体式方式集成分析过程的几个单元操作（预处理、分离、（生物）化学反应和检测阶段）。此外，由于其与生物材料的兼容性，LTCC 适用于基于酶反应的分析应用。在这里，我们展示了一种 LTCC 微流控系统的设计、构建和评估，该系统集成了一个微反应器（内部体积 24.28 μL），其中含有固定化的来自大肠杆菌的β-半乳糖苷酶（0.479 个活性单位）和一个光学流通池，以测量酶反应的产物。该酶固定在乙二醛琼脂糖载体上，在研究期间保持其活性。作为概念验证，通过测量通常用于活性测定的底物邻硝基苯-β-D-半乳糖吡喃糖苷的转化，对 LTCC-β-半乳糖苷酶系统进行了测试。一旦在整体集成的微柱中填充，就对微型化的流动系统进行了表征，优化了操作条件（流速和进样体积），并通过在毫摩尔水平上测定β-半乳糖苷酶的底物浓度成功地评估了其性能。
