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Analyzing antibody specificity with whole proteome microarrays
Gregory A Michaud1, Michael Salcius, Fang Zhou
1Protometrix, 688 East Main Street, Branford, Connecticut 06405, USA.
Nature Biotechnology
|November 11, 2003
Summary
Proteome arrays enable screening of antibody specificity against thousands of proteins. This study found that antibodies often cross-react with non-target proteins, highlighting the need for improved antibody design and selection methods.
Area of Science:
- Proteomics
- Immunology
- Biotechnology
Background:
- Commercial antibody availability is limited for many proteins.
- Increasing demand for specific antibodies in applications like antibody arrays and therapeutics necessitates reduced cross-reactivity.
- Current methods struggle to predict antibody cross-reactivity.
Purpose of the Study:
- To evaluate the utility of proteome arrays for assessing antibody specificity.
- To identify cross-reactivity issues with existing antibody reagents.
- To explore potential improvements in antibody design and selection.
Main Methods:
- Screening of 11 polyclonal and monoclonal antibodies against approximately 5,000 yeast proteins using a proteome array.
- Analysis of antibody binding to cognate and non-cognate proteins on the array.
- Sequence alignment to investigate potential reasons for cross-reactivity.
Main Results:
- Antibodies exhibited cross-reactivity with non-cognate proteins beyond their intended targets.
- The degree of cross-reactivity varied among the tested antibodies.
- Some cross-reactivity could be inferred from sequence similarity, but not predicted beforehand.
Conclusions:
- Proteome array technology is a valuable tool for assessing antibody specificity.
- The study revealed significant cross-reactivity issues with current antibodies.
- Proteome arrays offer potential for improving antibody design and selection for research and therapeutic applications.