My protein of interest is secreted in its mature form. I don't want to lyse the cells as the protocol indicates, because I don't want to purify any immature form of the protein. Is there a good way to use the superflow cartridge on cell media?
Many thanks!
Hi Christine,
The answer is likely yes, it is possible, but you should read Qiagen's technical documentation before trying it to optimize your chances of making it work. You may need to dilute your media out with specific buffers to ensure your solution that you load on the column does not interfere with the 6x-His binding to the Ni-NTA material. Cell culture media has various compounds, such as proteins from calf serum and added amino acids such as glutamine, which can interfere with this binding reaction and prevent the purification from working as intended.
Take a look at Qiagen's technical documentation (see page 74 of the document found at the link below), you can find a list of reagents compatible with the Ni-NTA matrix.
https://www.qiagen.com/us/resources/resourcedetail?id=79ca2f7d-42fe-4d62-8676-4cfa948c9435&lang=en
Your exact question is addressed on pages 76-77, with suggestions of how to improve the binding if this is an issue. It appears that various people have done this successfully, with perhaps lower recovery rates than with lysed cells but it should be possible.
Good luck!
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