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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
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

Abstract 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.

Keywords

  • Immunology in the medical area
  • scFv
  • antibody microarrays
  • antibody chips
  • sensitivity
  • affinity-tag

Other

8th International Conference on Miniaturized Systems for Chemistry and Life Sciences
Published
  • ISSN: 0260-6291
  • ISBN: 9780854048960
Carl B
Carl Borrebaeck
E-mail: carl.borrebaeck [at] immun.lth.se

Professor

Department of Immunotechnology

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