Peptide-tags for enhanced DNA microarray performance

DNA microarrays are powerful tools for gene expression analysis and genotyping studies in research and diagnostic applications. A high sensitivity and short time-to-result are prerequisites for their practical application in the clinic. The hybridization efficiency of DNA microarrays depends on the...

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Bibliographic Details
Published in:Faraday discussions 2011, Vol.149, p.201-210
Main Authors: Schulze, Holger, Ross, Alan J, Ember, Stuart W J, Luby, Julie, Khondoker, Mizanur, Giraud, Gerard, Ciani, Ilenia, Tlili, Chaker, Papale, Davide, Terry, Jonathan G, Mount, Andrew R, Walton, Anthony J, Crain, Jason, Ghazal, Peter, Bachmann, Till T, Campbell, Colin J
Format: Article
Language:English
Subjects:
Amino acid composition
Amino groups
Aromatics
DNA microarrays
Gene expression
Genotyping
Hepacivirus - genetics
Hepatitis C, Chronic - virology
Histidine - chemistry
Humans
Immobilization
Nucleic Acid Hybridization - genetics
Nucleic Acid Hybridization - methods
Oligonucleotide Array Sequence Analysis - methods
Oligonucleotide Probes - chemistry
Oligonucleotides
Oligopeptides - chemistry
Probes
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Summary:DNA microarrays are powerful tools for gene expression analysis and genotyping studies in research and diagnostic applications. A high sensitivity and short time-to-result are prerequisites for their practical application in the clinic. The hybridization efficiency of DNA microarrays depends on the probe density and the probe orientation and thus their accessibility for target molecules. In order to find an optimal probe immobilization procedure a set of different oligonucleotide modifications was tested on epoxy silane functionalized glass slides. It was found that histidine-tagged oligonucleotides resulted in the highest amount of bound probe and by far the best hybridization efficiencies. The detection limit obtained with histidine-tagged probes was up to two orders of magnitude lower compared to commonly used probe modifications. In order to further investigate the binding mechanism of histidine-tags towards functionalized glass substrates a set of different peptide-tags with and without free terminal amino-groups and with different amino acid compositions was tested. The results indicate an impact of the terminal amino group on the covalent surface binding and of aromatic amino acid residues on the enhanced hybridisation efficiency.
ISSN:1359-6640
1364-5498
DOI:10.1039/c005491g