Protein Purification Made Easy

Global Life Sciences Solutions
Friday, 01 January, 2021


Protein Purification Made Easy

Summer is here and it is time for some fun in the sun — but when the sun is not shining, why not consider boosting your knowledge and skills these holidays as well?

Proteins can be isolated based on differences in their physical and chemical properties, and can be analysed and purified using techniques like centrifugation, electrophoresis and chromatography — all of which have different benefits and outcomes.

Tagged recombinant proteins are relatively straightforward to purify. We recommend researchers use an affinity resin that corresponds to the tag system used and the natural conditions for the protein to avoid precipitation and degradation. Two-step purification is commonly used, the first step being the use of an affinity chromatography resin and the second step using a size-exclusion chromatography resin. If your requirement for purity is high, add an additional intermediate step of ion exchange or hydrophobic interaction chromatography to increase purity. However, try to use as few steps as possible as adding steps will decrease overall protein yield.

After purification, you may need to cleave off the tag before using the protein. The tag-free target protein needs to be separated from the tag and target protein still containing the tag. On-column cleavage can also be used where protease is added to the column with the target protein bound, enabling cleavage and removal to be performed in one step.

For non-tagged proteins, more effort is required to develop the protocols. Experimental planning is important to make conscious decisions for what conditions to test, what to optimise, and the requirements for purity and yield. In our experience, successful protein purification requires following a multi-step approach to perform Capture, Intermediate Purification and Polishing (CiPP).

To control your protein purification and deliver significantly more information on your target protein and impurities we recommend the use of a protein purification system from our ÄKTA system range. If you are new to protein purification or looking for tips on how to run and take care of your ÄKTA system, our Cytiva team has put together a set of free online training modules. Learn more at cytiva.com.

Related Sponsored Contents

Phenom Pharos G2 Desktop FEG-STEM for in-house biological and materials science sample analysis

The highly automated Desktop Thermo Scientific Phenom Pharos G2 FEG-SEM with the new scanning...

Is MMS the key to RNA analytics?

MMS provides a comprehensive understanding of binding affinity and structural changes, offering...

Multi-Component FTIR Gas Analyser for Continuous Measurement of ppm to % vol in a Portable or Mobile Form

Protea's atmosFIRt transportable multigas analyser system represents one of the most...


  • All content Copyright © 2024 Westwick-Farrow Pty Ltd