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Design of recombinant antibody microarrays for serum protein profiling: Targeting of complement proteins

Author:
  • Johan Ingvarsson
  • Anette Larsson
  • Anders Sjöholm
  • Lennart Truedsson
  • Bo Jansson
  • Carl Borrebaeck
  • Christer Wingren
Publishing year: 2007
Language: English
Pages: 3527-3536
Publication/Series: Journal of Proteome Research
Volume: 6
Issue: 9
Document type: Journal article
Publisher: The American Chemical Society

Abstract english

Antibody-based microarrays is a novel technology with great promise for high-throughput proteomics. The process of designing high-performing arrays has, however, turned out to be challenging. Here, we have designed the next generation of a human recombinant scFv antibody microarray platform for protein expression profiling of nonfractionated biotinylated human plasma and serum proteomes. The setup, based on black polymer Maxisorb slides interfaced with a fluorescent-based read-out system, was found to provide specific, sensitive (subpicomolar (pM) range) and reproducible means for protein profiling. Further, a chip-to-chip normalization protocol critical for comparing data generated on different chips was devised. Finally, the microarray data were found to correlate well with clinical laboratory data obtained using conventional methods, as demonstrated for a set of medium abundant (micromolar (mu M) to nanomolar (nM) range) protein analytes in serum and plasma samples derived from healthy and complement-deficient individuals.

Keywords

  • Microbiology in the medical area
  • Immunology in the medical area
  • profiling
  • protein expression
  • recombinant antibody microarrays
  • serum proteomics
  • complement proteins

Other

Published
  • CREATE Health
  • ISSN: 1535-3893
Carl Borrebaeck
E-mail: carl [dot] borrebaeck [at] immun [dot] lth [dot] se

Professor

Department of Immunotechnology

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Director

Create Health

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