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          Institute: MPI für molekulare Genetik     Collection: Department of Vertebrate Genomics     Display Documents



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ID: 268500.0, MPI für molekulare Genetik / Department of Vertebrate Genomics
Subnanoliter enzymatic assays on microarrays
Authors:Angenendt, Philipp; Lehrach, Hans; Kreutzberger, Jürgen; Glökler, Jörn
Language:English
Date of Publication (YYYY-MM-DD):2005-02-08
Title of Journal:PROTEOMICS
Volume:5
Issue / Number:2
Start Page:420
End Page:425
Copyright:© 1999-2006 John Wiley & Sons, Inc.
Review Status:not specified
Audience:Experts Only
Abstract / Description: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.
Free Keywords:Drug screening; Enzymatic assay; Metagenomics; Multiple spotting technique; Protein microarray
Comment of the Author/Creator:Email: Philipp Angenendt (p.angenendt@dkfz-heidelberg.de)
External Publication Status:published
Document Type:Article
Version Comment:Automatic journal name synchronization
Communicated by:Hans Lehrach
Affiliations:MPI für molekulare Genetik
External Affiliations:DKFZ, Functional Genome Analysis, Im Neuenheimer Feld 580, D-69120 Heidelberg
Identifiers:ISSN:1615-9853
DOI:10.1002/pmic.200400955
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