Abstract
We developed an in vitro protein expression and interaction analysis platform based on a highly parallel and sensitive microfluidic affinity assay, and used it for 14,792 on-chip experiments, which exhaustively measured the protein-protein interactions of 43 Streptococcus pneumoniae proteins in quadruplicate. The resulting network of 157 interactions was denser than expected based on known networks. Analysis of the network revealed previously undescribed physical interactions among members of some biochemical pathways.
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Acknowledgements
We thank members of the Stanford microfluidics foundry for help with device fabrication. This work was supported in part by the US National Institutes of Health Director's Pioneer award (to S.R.Q.) and a Fulbright award (to D.G.).
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S.R.Q. and S.J.M. are authors on a pending patent based on the results of this research.
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Supplementary Figures 1–4, Supplementary Tables 1–4, Supplementary Methods (PDF 826 kb)
Supplementary Data
Protein interaction list in MIMIx format (XLS 33 kb)
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Gerber, D., Maerkl, S. & Quake, S. An in vitro microfluidic approach to generating protein-interaction networks. Nat Methods 6, 71–74 (2009). https://doi.org/10.1038/nmeth.1289
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DOI: https://doi.org/10.1038/nmeth.1289
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