Abstract
Affinity proteomics, represented by antibody arrays, is a multiplex technology for high-throughput protein expression profiling of crude proteomes in a highly specific, sensitive, and miniaturized manner. The antibodies are individually deposited in an ordered pattern, an array, onto a solid support. Next, the sample is added, and any specifically bound proteins are detected and quantified using mainly fluorescence as the mode of detection. The binding pattern is then converted into a relative protein expression map, or protein atlas, delineating the composition of the sample at the molecular level. The technology provides unique opportunities for various applications, such as protein expression profiling, biomarker discovery, disease diagnostics, prognostics, evidence-based therapy selection, and disease monitoring. Here, we describe the generation and use of planar antibody arrays for serum protein profiling.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Borrebaeck CAK, Wingren C (2009) Design of high-density antibody microarrays for disease proteomics: key technological issues. J Proteomics 72:928–935
Haab BB (2006) Applications of antibody array platforms. Curr Opin Biotechnol 17(4):415–421
Sanchez-Carbayo M (2011) Antibody microarrays as tools for biomarker discovery. Methods Mol Biol 785:159–182
Borrebaeck CAK, Wingren C (2011) Recombinant antibodies for the generation of antibody arrays. Methods Mol Biol 785:247–262
Borrebaeck CAK, Wingren C (2007) High-throughput proteomics using antibody microarrays: an update. Expert Rev Mol Diagn 7:673–686
Wingren C, Borrebaeck CAK (2007) Progress in miniaturization of protein arrays—a step closer to high-density nanoarrays. Drug Discov Today 12:813–819
Wingren C, Borrebaeck CAK (2008) Antibody microarray analysis of directly labelled complex proteomes. Curr Opin Biotechnol 19:55–61
Haab BB (2005) Antibody arrays in cancer research. Mol Cell Proteomics 4:377–383
Hanash S (2003) Disease proteomics. Nature 422:226–232
MacBeath G (2002) Protein microarrays and proteomics. Nat Genet 32 Suppl:526–532
Carlsson A, Wuttge DM, Ingvarsson J, Bengtsson AA, Sturfelt G, Borrebaeck CAK, Wingren C (2011) Serum protein profiling of systemic lupus erythematosus and systemic sclerosis using recombinant antibody microarrays. Mol Cell Proteomics 10:M110 005033
Haab BB, Dunham MJ, Brown PO (2001) Protein microarrays for highly parallel detection and quantitation of specific proteins and antibodies in complex solutions. Genome Biol 2:RESEARCH0004
Wingren C, Ingvarsson J, Dexlin L, Szul D, Borrebaeck CAK (2007) Design of recombinant antibody microarrays for complex proteome analysis: choice of sample labeling-tag and solid support. Proteomics 7:3055–3065
Ingvarsson J, Larsson A, Sjoholm AG, Truedsson L, Jansson B, Borrebaeck CAK, Wingren C (2007) Design of recombinant antibody microarrays for serum protein profiling: targeting of complement proteins. J Proteome Res 6:3527–3536
Acknowledgements
This work was supported by grants from the Swedish National Science Council (VR-NT and VR-M), the SSF Strategic Center for Translational Cancer Research (CREATE Health), and Vinnova.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media, New York
About this protocol
Cite this protocol
Borrebaeck, C.A.K., Wingren, C. (2014). Antibody Array Generation and Use. In: Ossipow, V., Fischer, N. (eds) Monoclonal Antibodies. Methods in Molecular Biology, vol 1131. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-992-5_36
Download citation
DOI: https://doi.org/10.1007/978-1-62703-992-5_36
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-991-8
Online ISBN: 978-1-62703-992-5
eBook Packages: Springer Protocols