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Updated: May 30, 2026
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Analysis of Histone Antibody Specificity with Peptide Microarrays
Published on: August 1, 2017
Antibody array analysis of labelled proteomes: how should we control specificity?
Anders Holm1, Weiwei Wu, Fridtjof Lund-Johansen
1Department of Immunology, Clinic of Specialized Medicine and Surgery Oslo University Hospital, Rikshospitalet, N-0027 Oslo, Norway.
New Biotechnology
|August 16, 2011
Summary
Researchers developed Size-MAP, an antibody array Western blot, to assess protein binder specificity in multiplexed assays. This method helps validate binders for proteome-wide antibody arrays, addressing concerns about specificity in complex biological samples.
Area of Science:
- Proteomics
- Biotechnology
- Analytical Chemistry
Background:
- Multiplexed assays using protein binders face challenges in achieving proteome-wide coverage and specificity.
- Skepticism exists regarding the reliability of immobilized protein binders and labeled samples for specific protein detection.
Purpose of the Study:
- To review evidence supporting antibody array analysis for meaningful data generation.
- To address skepticism about specificity in antibody array-based detection.
- To introduce and validate a novel assay, Size-MAP, for assessing protein binder specificity.
Main Methods:
- Developed Size-MAP (size exclusion chromatography-resolved microsphere-based affinity proteomics).
- Combined size exclusion chromatography (SEC) for protein separation with microsphere-based antibody arrays.
- Utilized labeled sample proteins separated by SEC prior to array interaction.
Main Results:
- Size-MAP functions as an 'antibody array Western blot', resolving binder reactivity against protein size.
- The assay effectively discriminates between monospecific and polyreactive antibodies.
- Enabled automatic detection of binders reacting with the same target protein.
Conclusions:
- Size-MAP provides a direct method to test antibody specificity within array-based measurements.
- This approach aids in selecting high-quality protein binders for proteomic applications.
- The findings contribute to determining the feasibility of proteome-wide DNA microarrays.
