title: Design of recombinant antibody microarrays for serum protein profiling: targeting of complement proteins.
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 (microM) to nanomolar (nM) range) protein analytes in serum and plasma samples derived from healthy and complement-deficient individuals.
authors: Department of Immunotechnology, BMC D13, Lund University, SE-221 84 Lund, Sweden.
authors: Johan
authors: J
authors: Ingvarsson
authors: Anette
authors: A
authors: Larsson
authors: Anders G
authors: AG
authors: Sjöholm
authors: Lennart
authors: L
authors: Truedsson
authors: Bo
authors: B
authors: Jansson
authors: Carl A K
authors: CA
authors: Borrebaeck
authors: Christer
authors: C
authors: Wingren
mesh_terms: D019921:Biotinylation; D001798:Blood Proteins / Q000737:chemistry; D003165:Complement System Proteins / Q000737:chemistry; D006801:Humans; D008956:Models, Chemical; D040081:Protein Array Analysis / Q000379:methods*; D021381:Protein Transport; D011506:Proteins / Q000737:chemistry; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D001798:Blood Proteins; D011506:Proteins; D020543:Proteome; D011994:Recombinant Proteins; D003165:Complement System Proteins