Edit
Protein quality control for downstream processes – Course and Conference Office

EMBL Course

Protein quality control for downstream processes

Overview

Course overview

Purified proteins are used in various types of scientific experiments and fields. For example, in structural biology insights into the functional mechanisms can be obtained by elucidating the 3D molecular structure of proteins and protein complexes using technologies such as X-ray crystallography, cryo-EM and NMR. In biochemistry and biophysics, interactions with other proteins, nucleic acids and small molecules can be studied by determining affinities and specificities. Proteins can function as antigens to generate specific antibodies or as reagents in cell biology experiments. Furthermore, recombinant proteins can be used as tool molecules in genomics, chemical biology and microscopy assays. In order for these experiments to produce reliable and biologically relevant results, they must be performed using high-quality proteins that are active, properly folded, in the right oligomeric state and contain correctly inserted co-factors. Ensuring that these parameters are fulfilled requires quality control.

In this course, we want to help the participants to become acquainted with the concept of protein quality control and provide them with an overview of the specific protein requirements for various downstream applications. Different techniques that can be applied for this purpose will be introduced and participants will have the opportunity to get hands-on experience with these technologies.

Audience

This course is aimed at life science researchers of all levels, who are using or plan to use purified proteins in their wet lab experiments, but who do not necessarily have a strong background in protein production and characterisation methods. We will focus on introducing protein quality control methodologies in order to improve the reliability and reproducibility of data obtained from experiments that make use of purified proteins. After this course, participants should be familiar with the biochemical and biophysical techniques that can be applied to ensure that they are working with high-quality protein samples suitable for the planned downstream experiments.

Modules/resources

Basic information

  • Protein expression and purification process
  • Protein concentration
  • Buffer conditions (pH, ionic strength, buffer components)

Purity and intactness

  • SDS-PAGE → right size, no degradation?
  • UV-VIS spectrum → nucleic acid contaminations?
  • Mass spectrometry → expected molecular weight and amino acid sequence?

Homogeneity/polydispersity

  • SEC, SEC-MALS → oligomeric state, no aggregates, molar mass?
  • DLS → presence of aggregates?
  • Mass photometry → oligomeric state, molar mass?
  • Flow Induced Dispersion Analysis → presence of aggregates?

Folding state and conformational stability:

  • Circular dichroism → secondary structure? influence of buffer conditions and/or ligands?
  • Nano-DSF, thermofluor → optimal buffer conditions? thermal stability?

Activity

  • Enzymatic assays → functionality?
  • Interaction studies: ITC, MST, Flow Induced Dispersion Analysis → affinity for binding partners?

Learning outcomes

During the course, specialists from both academia and industry will work closely together to introduce technologies frequently used in protein quality control. Some of these methodologies can be utilised both to assess protein quality and as standalone biophysical characterisation techniques.

Our main goals are:

  • Teach the participants how to perform proper protein quality control checks
  • Introduce the participants to various biophysical techniques used for protein characterization, such as SEC-MALS, DLS, SLS, mass photometry, nano-DSF, flow induced dispersion analysis and spectroscopy
  • Show participants how to characterize protein interactions using ITC, MST and flow induced dispersion analysis
  • Where possible, allow participants to apply these technologies onto their own protein samples in the practicals
  • Overall, increase the success rate, reliability and reproducibility of downstream experiments in which purified proteins are used

What past participants say about the course

“This advanced course was my first “live” experience at the EMBL and it was superb. It was packed with great lectures, state-of-the-art technology, fruitful discussions, and beautiful people. The course was also a great opportunity to meet new people and expand your network. Highly recommend and I will definitely attend the next event.” – Klemen Mezgec, National Institute of Chemistry Ljubljana, Slovenia

Date: 19 - 22 Mar 2024

Location: EMBL Heidelberg

Venue: EMBL Advanced Training Centre


Deadline(s):

Registration: Closed


Organisers:


EMBL Courses and Conferences are kindly supported by our Corporate Partnership Programme

Corporate partners










































Associate partners

Edit

What's new on our blog

Edit