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

Författare:
Publiceringsår: 2007
Språk: Engelska
Sidor: 3527-3536
Publikation/Tidskrift/Serie: Journal of Proteome Research
Volym: 6
Nummer: 9
Dokumenttyp: Artikel
Förlag: American Chemical Society

Sammanfattning

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.

Disputation

Nyckelord

  • Medicine and Health Sciences
  • profiling
  • protein expression
  • recombinant antibody microarrays
  • serum proteomics
  • complement proteins

Övriga

Published
Yes
  • ISSN: 1535-3893

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