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



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ID: 126672.0, MPI für molekulare Genetik / Department of Vertebrate Genomics
3D protein microarrays: Performing multiplex immunoassays on a single chip
Authors:Angenendt, Philip; Glokler, Jörn; Konthur, Zoltán; Lehrach, Hans; Cahill, Dolores J.
Language:English
Date of Publication (YYYY-MM-DD):2003-09-01
Title of Journal:Analytical Chemistry
Journal Abbrev.:Anal. Chem.
Volume:75
Issue / Number:17
Start Page:4368
End Page:4372
Copyright:Copyright © 2003 American Chemical Society
Review Status:not specified
Audience:Experts Only
Abstract / Description:The enzyme-linked immunosorbent assay (ELISA) is typically applied in the format of microtiter plates. To increase throughput and reduce consumption of precious samples, efforts have been made to transfer ELISA to the microchip format using conventional microarrays, microfluidic systems, and chips bearing microwells. However, all three formats lack the possibility to screen several analytes on several immobilized binders at a time or require complicated liquid handling, surface modifications, and additional equipment. Here, we describe an immunoassay performed on a standard microscope slide without the requirement for wells or tubes to separate the samples using standard surfaces and machinery already available for microarray technology. The new multiple spotting technique (MIST) comprises immobilization of a binder onto a surface and subsequent spotting of the second compound on the same spot, on top of the immobilized binder. We show that the analytes bind their ligands immediately within the confined space of separate droplets on the chip surface, thereby eliminating the need for extra incubation time. We illustrate the feasibility of the new technique by spotting dilution rows of proteins or monoclonal and polyclonal antibodies on top of their immobilized binders. Moreover, we demonstrate specificity by applying a mixture of antibodies in a multiplex format and demonstrate that the technique is compatible with conventional microarray protocols, such as total incubation. Finally, we indicate that the technique is capable of quantifying as little as 400 zmol (240 000 molecules) of analyte.
Comment of the Author/Creator:Date: 2003, SEP 1
External Publication Status:published
Document Type:Article
Version Comment:Automatic journal name synchronization
Communicated by:Hans Lehrach
Affiliations:MPI für molekulare Genetik
External Affiliations:Max Planck Inst Mol Genet, D-14195 Berlin, Germany.; Royal Coll Surgeons Ireland, Natl Ctr Human Proteom, Dublin 2, Ireland.; PORTAGEN, D-44227 Dortmund, Germany.
Identifiers:ISI:000185192300011 [ID No:1]
ISSN:0003-2700 [ID No:2]
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