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

Forskningsoutput: TidskriftsbidragArtikel i vetenskaplig tidskrift

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.

Detaljer

Författare
Enheter & grupper
Forskningsområden

Ämnesklassifikation (UKÄ) – OBLIGATORISK

  • Mikrobiologi inom det medicinska området
  • Immunologi inom det medicinska området

Nyckelord

Originalspråkengelska
Sidor (från-till)3527-3536
TidskriftJournal of Proteome Research
Volym6
Utgivningsnummer9
StatusPublished - 2007
PublikationskategoriForskning
Peer review utfördJa