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Scientific_Methods t1_iraxe39 wrote

Reply to comment by Joe_Q in is it possible to synthesize proteins chemically? by yeeturking

To add to this. Many proteins require post-translational modifications that bacteria cannot perform. In those cases yeast cells are often used to produce the protein.

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Alwayssunnyinarizona t1_iraz0ro wrote

Or insect, or mammalian, depending on the application.

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Mikedc1 t1_irb31kf wrote

Then downstream, depending on its properties it may be difficult to separate it from contaminants from the upstream processes mentioned above. Attachment to some resin based on charge or a binding site can be used in a chromatography setup. Solubility or inertness in some chemical can help. Finally regular filtration or some sort of hollow fibre tff setup.

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NosemaCeranae t1_irbbvy9 wrote

> Attachment to some resin based on charge or a binding site can be used in a chromatography setup. Solubility or inertness in some chemical can help. Finally regular filtration or some sort of hollow fibre tff setup.

Yep. A very typical workflow would be preliminary semi-purification over an affinity column followed by isolation of your protein of interest over a size exclusion column. Just one, but very common, method.

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Jordanno99 t1_irdxll2 wrote

You normally add a tag such as ctag, histag, monoFc or biotin to assist with purification.

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