Protein Microarrays for Gene Expression and Antibody Screening

Proteins translate genomic sequence information into function, enabling biological processes. As a complementary approach to gene expression profiling on cDNA microarrays, we have developed a technique for high-throughput gene expression and antibody screening on chip-size protein microarrays. Using...

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Bibliographic Details
Published in:	Analytical biochemistry Vol. 270; no. 1; pp. 103 - 111
Main Authors:	Lueking, Angelika, Horn, Martin, Eickhoff, Holger, Büssow, Konrad, Lehrach, Hans, Walter, Gerald
Format:	Journal Article
Language:	English
Published:	United States Elsevier Inc 15-05-1999
Subjects:	antibody Antibody Specificity DNA, Complementary Escherichia coli expression Filtration gene Gene Expression Humans Ligands microarray Microchemistry - methods protein Proteins - genetics Proteins - immunology robotics screening Sensitivity and Specificity microarray screening expression antibody gene protein robotics
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Summary:	Proteins translate genomic sequence information into function, enabling biological processes. As a complementary approach to gene expression profiling on cDNA microarrays, we have developed a technique for high-throughput gene expression and antibody screening on chip-size protein microarrays. Using a picking/spotting robot equipped with a new transfer stamp, protein solutions were gridded onto polyvinylidene difluoride filters at high density. Specific purified protein was detected on the filters with high sensitivity (250 amol or 10 pg of a test protein). On a microarray made from bacterial lysates of 92 human cDNA clones expressed in a microtiter plate, putative protein expressors could be reliably identified. The rate of false-positive clones, expressing proteins in incorrect reading frames, was low. Product specificity of selected clones was confirmed on identical microarrays using monoclonal antibodies. Cross-reactivities of some antibodies with unrelated proteins imply the use of protein microarrays for antibody specificity screening against whole libraries of proteins. Because this application would not be restricted to antigen–antibody systems, protein microarrays should provide a general resource for high-throughput screens of gene expression and receptor–ligand interactions.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0003-2697 1096-0309
DOI:	10.1006/abio.1999.4063