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Subnanoliter enzymatic assays on microarrays
Author(s) -
Angenendt Philipp,
Lehrach Hans,
Kreutzberger Jürgen,
Glökler Jörn
Publication year - 2005
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200400955
Subject(s) - multiplex , microtiter plate , dna microarray , microarray , computational biology , biology , high throughput screening , microbiology and biotechnology , biochemistry , bioinformatics , gene , gene expression
Many areas of research today are based on enzymatic assays most of which are still performed as enzyme‐linked immunosorbent assays in microtiter plates. The demand for highly parallel screening of thousands of samples eventually led to a miniaturization and automation of these assays. However, the final transfer of enzymatic assays from a microtiter‐based technology to microarrays has proven to be difficult for various reasons, such as the inability to maintain unbound reaction products on the spot of reaction or the missing capability of multiplexing. Here, we have conducted multiplex enzymatic assays in subnanoliter volumes on a single microarray using the multiple spotting technology. We were able to measure enzymatic activity with a sensitivity down to 35 enzyme molecules, applying only conventional flat microarray surfaces and standard microarray hardware. We have performed assays of inhibition and applied this format for the detection of prognostic markers, such as cathepsin D. The new approach allows the rapid and multiplex screening of thousands of samples on a single microarray with applications in drug screening, metagenomics, and high‐throughput enzyme assays.