Antibody Microarrays

Christer Wingren, Lund University, Lund, Sweden
Carl Borrebaeck, Lund University, Lund, Sweden

Published online: January 2006

DOI: 10.1038/npg.els.0004026

Antibody-based microarray is a novel technology with great promise within proteomics. A microarray can be printed with thousands of antibodies carrying the desired specificities, the biological sample added (e.g. a proteome) and any specifically bound analytes detected. Global proteome analysis and protein expression profiling using this tool will provide new opportunities for biomarker discovery, drug target identification, disease diagnostics and insights into disease biology.

Keywords: antibody microarrays; antibody chips; protein microarrays; probes; antibodies; proteomics

Figure 1. Schematic illustration of the antibody microarray concept.
Figure 2. Schematic illustration of a scFv antibody library.
Figure 3. Schematic illustration of the three major probe-binding approaches currently used; (a) adsorption, (b) covalent binding, and (c) affinity binding.
Figure 4. Comparative proteome analysis (cell lysate) of nonstimulated versus stimulated dendritic cells using a nine probe anti-cytokine antibody microarrays. (a) Cy5-scanned microarray image of activated sample. (b) Cy3-scanned microarray image of nonactivated sample. (c) Comparison of cytokine levels in activated versus nonactivated sample. Four cytokines were found to be up-regulated in the activated sample. Data adopted from Wingren et al. 2003.
close
 References
    Borrebaeck CAK (2000) Antibodies in diagnostics – from immunoassays to protein chips. Immunology Today 21: 379–382.
    Haab BB, Dunham MJ and Brown PO (2000) Protein microarrays for highly parallel detection and quantification of specific proteins and antibodies in complex solutions. Genomebiology 1: 1–22.
    Knezevic V, Leethanakul C, Bichsel VE et al. (2001) Proteomic profiling of the cancer microenvironment by antibody arrays. Proteomics 1: 1271–1278.
    Kusnezow W and Hoheisel JD (2002) Antibody microarrays: promises and problems. Biotechniques 33: S14–S23.
    Lin Y, Huang R, Chen LP et al. (2003) Profiling of cytokine expression by biotin-labeled based protein arrays. Proteomics 3: 1750–1757.
    MacBeath G (2002) Protein microarrays and proteomics. Nature Genetics 32: 526–532.
    MacBeath G and Schreiber SL (2000) Printing proteins as microarrays for high-throughput function determination. Science 289: 1760–1763.
    Miller JC, Zhou H, Kwekel J et al. (2003) Antibody microarray profiling of human prostate cancer sera: antibody screening and identification of potential biomarkers. Proteomics 3: 56–63.
    Sreekumar A, Nyati MK, Varambally S et al. (2001) Profiling of cancer cells using protein microarrays: discovery of novel radiation-regulated proteins. Cancer Research 61: 7585–7593.
    Steinhauer C, Wingren C, Malmborg-Hager A and Borrebaeck CAK (2002) Single framework recombinant antibody fragments designed for protein chip applications. Biotechniques 33: S38–S45.
    Söderlind E, Strandberg L, Jirholt P et al. (2000) Recombining germline-derived CDR sequences for creating diverse single-framework antibody libraries. Nature Biotechnology 18: 852–856.
    Wilson DS and Nock S (2003) Recent developments in protein microarray technology. Angewandte Chemie 42: 494–500.
    Wingren C, Ingvarsson J, Lindstedt M and Borrebaeck CAK (2003) Recombinant antibody microarrays – a viable option? Nature Biotechnology 21: 223.
    book Wingren C, Montelius L and Borrebaeck CAK (2004) "Mega-dense nanoarrays – the challenge of novel antibody array formats". In: Schena M and Weaver S (eds), Protein Microarrays, "Chap. 17". pp. 339–352. Sudbury, MA: Jones and Bartlett.
    Wingren C, Steinhauer C, Ingvarsson J et al. (2005) Microarrays based on affinity-tagged scFv antibodies: sensitive detection of analyte in complex proteomes. Proteomics 5(5): 1281–1291.
    Zhu H and Snyder M (2003) Protein chip technology. Current Opinion in Chemical Biology 7: 55–63.
 Further Reading
    Borrebaeck CAK, Ekström S, Malmborg-Hager AC et al. (2001) Protein chips based on recombinant antibody fragments: a highly sensitive approach as detected by mass spectrometry. Biotechniques 30: 1126–1132.
    Haab B (2003) Methods and applications of antibody microarrays in cancer research. Proteomics 3: 2116–2122.
    Kingsmore SF and Patel DD (2003) Multiplexed protein profiling in antibody-based microarrays by rolling circle amplification. Current Opinion in Biotechnology 14: 74–81.
    Peluso P, Wilson DS and Do D (2003) Optimizing antibody immobilization strategies for the construction of protein microarrays. Analytical Biochemistry 312: 113–124.
    Svedhem S, Pfeiffer I, Larsson C et al. (2003) Patterns of DNA-labelled and protein/scFv-carrying lipid vesicles directed by preferential protein adsorption and supported lipid bilayer formation on an Au/SiO2 template. Chem Biol Chem 4: 339–343.
    Zhou H, Bouwman K, Schotanus M et al. (2004) Two-color, rolling circle amplification on antibody microarrays for sensitive multiplexed serum-protein measurements. Genome Biology 5: R28.
    Belov L, de la Vega O, dos Remedios CG, Mulligan SP and Christopherson RI (2001) Immunophenotyping of leukemias using a cluster of differentiation antibody microarray. Cancer Research 61: 4483–4489.
    Pawlak M, Schick E, Bopp MA et al. (2003) Zeptosen's protein microarrays: a novel high performance microarray platform for low abundance protein analysis. Proteomics 2: 383–393.
    Pavlickova P, Knappik A, Kambhampati D, Ortigao F and Hug H (2003) Microarray of recombinant antibodies using a streptavidin sensor surface self-assembled onto a gold layer. Biotechniques 34: 124–130.

Related Sub-Topics:

Diagnostics, Therapeutics and Methods | Techniques and Tools in Molecular Biology | Biochemical and Biophysical Techniques
Contact Editor close
Submit a note to the editor about this article by filling in the form below.

* Required Field

Article Title: Antibody Microarrays
 
How to Cite close
Wingren, Christer, and Borrebaeck, Carl(Jan 2006) Antibody Microarrays. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0004026]