Can I purify protein straight from the cell culture supernatant?
It depends on the purification techniques. As far as we are aware, you can use techniques such as ion exchange or antibody affinity purification. However, Ni-NTA purification or other metal-affinity purification of His-tagged proteins will not work. Firstly, the cell culture medium contains other charged compounds that will compete with your protein. Secondly, the low pH (~5.5) causes the His-tag to be protonated and therefore precludes binding to the resin. You will have to change buffer to a higher pH buffer (pH 7.5+) to deprotonate the His-tag. See us on how to do this using a tangential flow filtration system and check out the protocols page.
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- Can I purify protein straight from the cell culture supernatant?