Science Webinar Series

Revealing the secrets of intractable cellular functions: All-in-one-well methods for studying protein interaction and secretion

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Science  24 Apr 2015:
Vol. 348, Issue 6233, pp. 461
DOI: 10.1126/science.348.6233.461-c

Summary

Protein-based assays are essential tools for many laboratories in both industrial and academic settings. Protein targets of interest are often found at low abundance and can be complex in nature, making their analysis intractable. One of the considerable challenges when developing assays is that small sample sizes can limit the amount of relevant information that can be extracted. The ability to miniaturize an assay allows sufficient data to be collected from scarce samples, while being able to interrogate multiple proteins simultaneously through multiplexing enables researchers to extract more data from each experiment and improve the biological relevance of the results significantly. This multiplexing ability provides the opportunity to quantitatively profile multiple signaling pathways from minute samples derived from very small cell populations. Furthermore, having the tools to directly study protein target binding in cells allows biologically relevant information to be preserved, including subcellular localization, posttranslational modifications, and the occurrence of multiple interactions with different accessory proteins and other scaffolding molecules. In this webinar, two examples of such approaches will be discussed. The first is the use of a method to amplify the signal in a target engagement assay using small samples, utilized to interrogate modulators of cytokine secretion. The second involves a so-called cellular thermal shift assay (CETSA) to detect protein-ligand interactions without the need to modify either the ligand or receptor, or the necessity for recombinant cell lines.