Design of recombinant antibody microarrays for serum protein profiling: Targeting of complement proteins
Författare
Summary, in 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.
Avdelning/ar
Publiceringsår
2007
Språk
Engelska
Sidor
3527-3536
Publikation/Tidskrift/Serie
Journal of Proteome Research
Volym
6
Issue
9
Länkar
Dokumenttyp
Artikel i tidskrift
Förlag
The American Chemical Society (ACS)
Ämne
- Microbiology in the medical area
- Immunology in the medical area
Nyckelord
- profiling
- protein expression
- recombinant antibody microarrays
- serum proteomics
- complement proteins
Status
Published
ISBN/ISSN/Övrigt
- ISSN: 1535-3893