Abstract
With the increasing availability of collections of antibodies, their evaluation in terms of binding selectivity becomes an important but challenging task. Planar antigen microarrays are very suitable tools to address this task and provide a powerful proteomics platform for the characterization of the binding selectivity of antibodies toward thousands of antigens in parallel. In this chapter, we describe our in-house developed procedures for the generation of high-density planar antigen microarrays with over 21,000 features. We also provide the details of the assay protocol, which we routinely use for the assessment of binding selectivity of the polyclonal antibodies generated within the Human Protein Atlas.
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Acknowledgments
We thank the entire staff of the Human Protein Atlas for producing all the antigens and antibodies utilized in this study. This work was supported by the ProNova VINN Excellence Centre for Protein Technology (VINNOVA, Swedish Governmental Agency for Innovation Systems) and by grants from the Knut and Alice Wallenberg Foundation, SciLifeLab Stockholm and the KTH Center for Applied Proteomics funded by the Erling-Persson Family Foundation. The authors declare no conflict of interest.
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Sjöberg, R. et al. (2018). High-Density Antigen Microarrays for the Assessment of Antibody Selectivity and Off-Target Binding. In: Rockberg, J., Nilvebrant, J. (eds) Epitope Mapping Protocols. Methods in Molecular Biology, vol 1785. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7841-0_15
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DOI: https://doi.org/10.1007/978-1-4939-7841-0_15
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