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Purified proteins are used in many biological experiments, including for structure determination, studying protein interactions, identifying drug metabolism and specificity, mechanistic studies, in vivo experiments and many other applications. In order for these experiments to be biologically relevant, they must be performed using high quality protein that is correctly folded, active, contains properly inserted cofactors and is in the correct oligomeric state. Ensuring that these parameters are fulfilled requires protein quality control. In this course, we will review some quality control basics and their application along with a description of various techniques that may be used to ensure high quality samples to produce reliable and reproducible data in downstream processes.
This course is aimed at life science researchers of all levels who would like to learn how to assess the quality of their purified proteins. Prior experience with protein quality control methods is not necessary, but it might be helpful to have a basic understanding of protein expression and purification. The course will focus on introducing the importance of protein quality control and how to approach and apply various techniques for ensuring high quality samples that will produce reliable data in downstream applications.
- Thermal shift assays: NanoDSF and Thermofluor for buffer screening
- Interaction studies: ITC and MST
- Light Scattering: SEC-MALS and DLS
- Cryo-EM: how sample quality control can make a difference in resolution
After this course you should be able to:
- Assess the quality of your protein(s) and protein complexes using the methods presented in the course
- Analyse and interpret the data obtained from different biochemical and biophysical methods commonly used in protein quality control
- Know which methods and technologies you can apply to try to improve the quality of your protein(s) and protein complexes
- Decide which quality control parameters are the most important ones for different types of downstream applications