Antibody Microarrays


Antibody‐based microarray is a recently established proteomic technology, providing unique opportunities for multiplexed protein expression profiling of crude, non‐fractionated samples in a specific, sensitive and miniaturised manner. A microarray can be printed with a few up to several thousands of different antibodies carrying the desired specificities, the biological sample added (e.g. a serum sample) and any specifically bound analytes detected and quantified. The obtained microarray images are then transformed into protein expression profiles, or protein maps, revealing the composition of the sample at the molecular level. Bioinformatics is used to handle the often large‐scale data set(s) acquired. Protein expression profiling using this tool will provide new opportunities for biomarker discovery, drug target identification, (early) disease diagnostics and prognostics as well as insights into disease biology.

Key Concepts

  • Antibody microarrays are based on dispensing antibodies one‐by‐one in an ordered pattern, an array, onto a solid support.
  • Antibody‐based microarray is a miniaturised set‐up for simultaneously measuring numerous proteins in minute amount of sample.
  • Antibody microarrays can be used to perform multiplexed protein expression profiling of crude, complex samples.
  • Using antibody microarrays, less than a drop of blood is required in order to generate a protein map, or protein fingerprint, reflecting the protein content in the sample.
  • Multiplexed protein expression profiling using antibody microarrays provides novel opportunities for biomarker discovery, disease diagnosis and prognosis.

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. Schematic illustration of antibody microarray‐based applications.


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Further Reading

Alhamdani MS, Schröder C and Hoheisel JD (2010) Analysis conditions for proteomic profiling of mammalian tissue and cell extracts with antibody microarrays. Proteomics 10: 3203–3207.

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Dexlin‐Mellby L, Sandtsröm A, Antberg L, et al. (2011) Design of recombiant antibody microarrays for membrane protein profiling of cell lysate and tissue extracts. Proteomics 11: 1550–1554.

Haab B (2003) Methods and applications of antibody microarrays in cancer research. Proteomics 3: 2116–2122.

He M, Stoevesandt O and Taussig MJ (2008) In situ synthesis of protein arrays. Current Opinion in Biotechnology 19: 4–9.

Kingsmore SF and Patel DD (2003) Multiplexed protein profiling in antibody‐based microarrays by rolling circle amplification. Current Opinion in Biotechnology 14: 74–81.

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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.

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Wingren, Christer, and Borrebaeck, Carl(Oct 2015) Antibody Microarrays. In: eLS. John Wiley & Sons Ltd, Chichester. [doi: 10.1002/9780470015902.a0004026.pub2]