High-throughput proteomics using antibody microarrays: an update

Abstract

Antibody-based microarrays are a rapidly emerging technology that has advanced from the first proof-of-concept studies to demanding serum protein profiling applications during recent years, displaying great promise within disease proteomics. Miniaturized micro- and nanoarrays can be fabricated with an almost infinite number of antibodies carrying the desired specificities. While consuming only minute amounts of reagents, multiplexed and ultrasensitive assays can be performed targeting high- as well as low-abundance analytes in complex nonfractionated proteomes. The microarray images generated can then be converted into protein expression profiles or protein atlases, revealing a detailed composition of the sample. The technology will provide unique opportunities for fields such as disease diagnostics, biomarker discovery, patient stratification, predicting disease recurrence and drug target discovery. This review describes an update of high-throughput proteomics, using antibody-based microarrays, focusing on key technological advances and novel applications that have emerged over the last 3 years.

Identification of Serum Biomarker Signatures Associated with Pancreatic Cancer

“…Briefly, 121 human recombinant single-chain Fv (scFv) antibodies, targeting 57 mainly immunoregulatory biomolecules, were used as probes. The specificity, affinity (nanomolar range), and on-chip functionality of the phagedisplay-derived scFvs (19) was ensured by using (i) stringent selection protocols (19), (ii) multiple clones ( 4) per target molecule, and (iii) an scFv library microarray adapted by molecular design (20,21). The planar antibody microarrays (array size; 160 Â 8, <2 cm 2 ) were prepared by dispensing the antibodies and controls one-by-one (330 pL/drop) with a noncontact spotter.…”

High-content affinity-based proteomics: unlocking protein biomarker discovery

“…In the ensuing 10 years, there has been a large effort to develop single antibody array technology. Recent examples include a similar two-color approach with larger spotted arrays (810 antibodies) [53] and lovely work from Carl Borrebaeck's laboratory in Sweden, who have published excellent reviews of the technology [54,55]. In a recent paper, Proteomics and diagnostics: let's get specific, again, we reviewed the central issue of specificity for scaling proteomic technologies to deliver on the promise of high analyte density, sensitivity, and specificity [12].…”

“…In a recent paper, Proteomics and diagnostics: let's get specific, again, we reviewed the central issue of specificity for scaling proteomic technologies to deliver on the promise of high analyte density, sensitivity, and specificity [12]. Arrays of hundreds of printed single antibodies (antibody-capture arrays) are currently commercially available, but are limited to the sensitivity and specificity of a single antibody and suffer from cross-reactivity [12,54,55]. It is not possible, even with the very best monoclonals antibodies, to reliably measure intended analytes with single-non-sandwich reagents.…”

Serum cytokine modulation after Staphylococcus hyicus infection in BALB/c mice

“…The three platforms for antibody array approaches include sandwich-based, label-based, and competition-based antibody arrays (Borrebaeck and Wingren, 2007). Antibody array technology has emerged as a promising technology for disease biomarker discovery.…”