Meeting the Challenge of Optimizing His-tag Protein Purification

Expressing proteins modified with a His-tag is a very powerful and common way to efficiently purify your target molecule. The His-tag provides effective capture using Immobilized Metal-ion Affinity Chromatography (IMAC). But, as usual, not much is simple and there are challenges in the form of tagged proteins not binding and eluting as expected.

The binding can be affected by many things, such as: 

• The expression system used (inclusion bodies, secretion, PTM, proteolytic cleavage)
• The length of the His-tag
• Differences in affinity for the particular metal ion bound to the chelator on the resin
• The chelator ligand itself
• Reducing agents and chelating components in the feed
• The buffer composition during binding and elution.

So, even if the purification technique is simple in itself, it can be very hard to know what is wrong when no (or very little) of your target proteins are collected in your test tube. Where do you start in terms of troubleshooting? What are the signs to look for to understand what may need to be changed? How do you tackle the challenge of optimizing your His-tag protein purification method?

In this webinar, our featured speaker will go through the most common challenges, give their best hints & tips for what to look out for and what you can do to solve them. She will go through some application examples and discuss methods of screening before a decision is made in regards to which set-up and purification method to move forward with.

There are also new purification products available that specifically address the issue with components used in more complex expressions system such as insect cells or other eukaryotic expression systems. These expression systems often contain components that can interfere with the binding of the protein, for example stripping the metal ions that are needed for the binding.

At the end of the webinar, attendees will have a good overview on how to meet the challenges of optimizing His-tag protein purification and the options and tools that are available to help improve yield.

Speaker

Anna Heijbel, Sr Global Product Manager, Bio-Works

After finishing studies focused on biochemistry and biology at the University of Uppsala, Anna’s whole working life has revolved around the purification and characterization of different kinds of proteins, peptides, mAbs, etc. She has experience in purifying cell products from natural sources and recombinant tagged/untagged proteins at the lab bench as an application specialist at different diagnostic, pharma and biotech companies. She also has over 10 years’ experience as a product manager for different biotech companies.

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