Ceramic microsystem incorporating a microreactor with immobilized biocatalyst for enzymatic spectrophotometric assays

Mireia Baeza, Carmen López, Julián Alonso, Josep López-Santín, Gregorio Álvaro

Research output: Contribution to journalArticleResearchpeer-review

29 Citations (Scopus)

Abstract

Low-temperature coflred 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 μL) with an immobilized β-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-βgalactosidase system was tested by measuring the conversion of ortlio-nitrophenyl β-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 β-galactosidase substrate concentration at the millimolar level. © 2010 American Chemical Society.
Original languageEnglish
Pages (from-to)1006-1011
JournalAnalytical Chemistry
Volume82
Issue number3
DOIs
Publication statusPublished - 1 Feb 2010

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