Methods engineering of biomolecular and cellular assays
Problem solving subject to constraints is the hallmark of engineering design (1). We adopt a design approach to invent new methods for interrogating cells and biomolecules. Our goal is to develop bioassays that are sensitive, quantitative, and as high-throughput and multiplex as possible. Most importantly, they should be reliable, generalizable, and shareable.
In the realm of signal transduction, we have developed and expanded high-throughput methods for profiling protein phosphatase activities (2, 3). These methods are now being applied to chronic viral infections along with a technique under development for a multiplex kinase activity assay. We also seek to extend or standardize established methods. For example, we have scaled quantitative real-time PCR to the entire human signaling receptome (4) and set standards for quantitative immunoblotting (5). Beyond empirical methods, we have completed a software platform for digital segmentation of brightfield images of 3D organoid cultures (to be submitted).
- Janes KA, Chandran PL, Ford RM, Lazzara MJ, Papin JA, Peirce SM, Saucerman JJ, Lauffenburger DA. (2017) An engineering design approach to systems biology. Integr Biol, 9, 574-83. [Link to Article]
- Shah M, Smolko CM, Kinicki S, Chapman ZD, Brautigan DL, Janes KA. (2017) Profiling subcellular phosphatase responses to coxsackievirus B3 infection of cardiomyocytes. Mol Cell Proteomics, 16, S244-S262. [Link to Article]
- Janes KA. (2015) An analysis of critical factors for quantitative immunoblotting. Sci Signal, 8, rs2 [Link to Article]
- Bose AK, Janes KA. (2013) A high-throughput assay for phosphoprotein-specific phosphatase activity in cellular extracts. Mol Cell Proteomics, 12, 797-806. [Link to Article]
- Kang BH*, Jensen KJ*, Hatch JA, Janes KA. (2013) Simultaneous profiling of 194 distinct receptor transcripts in human cells. Sci Signal, 6, rs13. [Link to Article]