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Analyzing antibody specificity with whole proteome microarrays

Nature Biotechnology volume 21, pages 1509–1512 (2003)Cite this article

Abstract

Although approximately 10,000 antibodies are available from commercial sources, antibody reagents are still unavailable for most proteins1. Furthermore, new applications such as antibody arrays2,3,4,5 and monoclonal antibody therapeutics6,7 have increased the demand for more specific antibodies to reduce cross-reactivity and side effects. An array containing every protein for the relevant organism represents the ideal format for an assay to test antibody specificity, because it allows the simultaneous screening of thousands of proteins for possible cross-reactivity. As an initial test of this approach, we screened 11 polyclonal and monoclonal antibodies to ∼5,000 different yeast proteins deposited on a glass slide and found that, in addition to recognizing their cognate proteins, the antibodies cross-reacted with other yeast proteins to varying degrees. Some of the interactions of the antibodies with noncognate proteins could be deduced by alignment of the primary amino acid sequences of the antigens and cross-reactive proteins; however, these interactions could not be predicted a priori. Our findings show that proteome array technology has potential to improve antibody design and selection for applications in both medicine and research.

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Figure 1: Fluorescent images of antibody probes of the yeast proteome microarray.
Figure 2: Analysis of anti-Hda1 binding to the yeast proteome microarray.
Figure 3: Analysis of anti-Pep12 binding to the yeast proteome microarray.

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Authors and Affiliations

  1. Protometrix, 688 East Main Street, Branford, 06405, Connecticut, USA

    Gregory A Michaud, Michael Salcius, Fang Zhou, Rhonda Bangham, Jaclyn Bonin, Hong Guo, Paul F Predki & Barry I Schweitzer

  2. Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, 06520-8103, Connecticut, USA

    Michael Snyder

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  1. Gregory A Michaud
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Correspondence to Barry I Schweitzer.

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The authors are employees of Protometrix.

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Michaud, G., Salcius, M., Zhou, F. et al. Analyzing antibody specificity with whole proteome microarrays. Nat Biotechnol 21, 1509–1512 (2003). https://doi.org/10.1038/nbt910

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