Accelerating BTK Inhibitor Discovery: From DNA to Functional Protein Characterization in Just 5 Days

This application note showcases how Nuclera’s eProtein Discovery™ system, combined with Cytiva’s Biacore™ SPR platform, overcomes major bottlenecks in kinase drug discovery by enabling rapid, high-yield production and kinetic analysis of Bruton’s Tyrosine Kinase (BTK) variants.

Using an integrated workflow that merges AlphaFold-guided construct design, automated digital microfluidics, Cell-free synthesis and real-time label-free interaction studies, researchers screened, scaled up, and characterized BTK-inhibitor interactions with unprecedented speed—achieving functionally characterized proteins in only five days.

In this application note, you'll discover:

• A breakthrough 5-day workflow for kinase characterization, from DNA constructs to Biacore™ kinetic assays.

• How eProtein Discovery™ enables expression of soluble, functional BTK, leveraging construct optimization, cell-free protein synthesis, and strategic use of solubility tags.

• The role of Avi-tag biotinylation and SPR capture in streamlining BTK interaction analysis with next-gen inhibitors like fenebrutinib and vecabrutinib.

• Real-world insights into inhibitor binding kinetics, including complex interaction profiles uncovered by Single-Cycle Kinetics (SCK) on Biacore™ 1S+.