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Microarrays based on affinity-tagged SCFV antibodies: Sensitive detection of analyte in complex proteomes

Author

  • Christer Wingren
  • Cornelia Steinhauer
  • Johan Ingvarsson
  • E Persson
  • Kristina Larsson
  • Carl Borrebaeck

Summary, in English

Antibody microarray is a novel technology that will allow us to efficiently perform global proteome analysis. Performing rapid and multiplexed analysis on complex proteomes, such as human serum, targeting also low-abundant analytes will, however, place high demands upon the microarray design with respect to logistics, specificity and sensitivity. In this study, we have shown that non-purified affinity-tagged scFv antibody fragments could be directly applied as probes thereby eliminating the need for any time-consuming pre-purification steps. Further, highly functional arrays were obtained providing an assay sensitivity in the pM to fM range. In fact, 300 zeptomole analyte (approx. 5,000 molecules) may be sufficient for detection. Moreover, highly complex proteomes could be analysed without impairing the specificity and sensitivity of the setups. This is a pre-requisite for the design of high-density antibody arrays applied in high-throughput proteomics.

Publishing year

2005

Language

English

Pages

264-266

Publication/Series

Micro Total Analysis Systems 2004, Vol 2

Issue

297

Document type

Conference paper

Publisher

Royal Society of Chemistry

Topic

  • Immunology in the medical area

Keywords

  • scFv
  • antibody microarrays
  • antibody chips
  • sensitivity
  • affinity-tag

Conference name

8th International Conference on Miniaturized Systems for Chemistry and Life Sciences

Conference date

2004-09-26 - 2004-09-30

Conference place

Malmo, Sweden

Status

Published

ISBN/ISSN/Other

  • ISSN: 0260-6291
  • ISBN: 9780854048960