Purification-free affinity and concentration measurement of membrane-protein targets

This webinar introduces an integrated preparative and analytical method that enables researchers to simultaneously measure:

• The concentration of an endogenous membrane-protein target
• Its binding affinity to a specific ligand in a native lipid-bilayer environment directly extracted from crude cellular membranes

To demonstrate this approach, Sebastian Fiedler and Sandro Keller will describe how the nanodisc-forming polymer, Glyco-DIBMA, was used to create a native membrane-protein library of a breast cancer cell line. They will discuss how microfluidic diffusional sizing (MDS) on the Fluidity One-M, was then used to determine both the concentration of the endogenous oncoprotein HER2 and its affinity to trastuzumab, a therapeutic HER2-specific antibody. The combination of native membrane-protein libraries with MDS provides quantitative information on membrane proteins without the need for laborious and costly protein-purification campaigns. The method is straightforward to adapt in standard research laboratories, takes only a few hours to complete, and has the potential to be expanded from cell lines to tissues and tumor biopsies.

Key learning objectives

• Advantages of measuring biomolecular interactions in solution by MDS and how to use this technology in measuring affinity and concentration of an unpurified membrane-protein target in a complex background
• Workflow and timings of a purification-free method for the extraction of integral membrane proteins from cellular membranes without disrupting their local lipid-bilayer environment by using native nanodiscs
• Case-study results on obtaining both the absolute cellular concentration and the antibody affinity of the HER2 oncoprotein embedded in native nanodiscs yielded by MDS technology

Who should attend?

Drug-discovery scientists and academic scientists working on membrane-protein targets

Certificate of attendance

All webinar participants can request a certificate of attendance, including a learning outcomes summary, for continuing education purposes.