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

Johan Ingvarsson, Anette Larsson, Anders Sjöholm, Lennart Truedsson, Bo Jansson, Carl Borrebaeck, Christer Wingren

Research output: Contribution to journalArticlepeer-review

Abstract

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.
Original languageEnglish
Pages (from-to)3527-3536
JournalJournal of Proteome Research
Volume6
Issue number9
DOIs
Publication statusPublished - 2007

Subject classification (UKÄ)

  • Microbiology in the medical area
  • Immunology in the medical area

Free keywords

  • profiling
  • protein expression
  • recombinant antibody microarrays
  • serum proteomics
  • complement proteins

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