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Competitive Assay Formats for High-Throughput Affinity Arrays

Tricia Diggle

Jonathan Terrett

Mikhail Soloviev

Oxford GlycoSciences (UK) Ltd., Abingdon, United Kingdom. mikhail.soloviev{at}ogs.co.uk

The authors describe a novel method for the quantitation of differential levels of biomolecules using unlabeled samples and protein-binding arrays for assessing differential expression. Traditional affinity arrays, whether in microplates or protein microarrays, suffer from a few common problems—a shortage of characterized antibodies and highly variable affinities for those available. Also, the assayed proteins could be present in a wide range of concentrations and physicochemical properties, so that it becomes an onerous task to optimize assay conditions for each antibody-antigen pair. Currently, this restricts parallel affinity assays to a low number of carefully selected antibodies and restricts the development of highly multiplexed parallel affinity assays. A displacement strategy allows the use of a much wider range of antibodies, reducing the requirement for matched affinities. The competitive assays described here also show a much higher tolerance for nonspecific background noise. The range of assayed protein concentrations is only limited by the sensitivity of the detection system used. (Journal of Biomolecular Screening 2003:257-263)

Key Words: affinity assay • protein array • displacement • competitive binding • multiplexing

Journal of Biomolecular Screening, Vol. 8, No. 3, 257-263 (2003)
DOI: 10.1177/1087057103008003003


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