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Refolding Insoluble Purified Proteins

Refolding Insoluble Purified Proteins - Recombinant Protein Forum

Refolding Insoluble Purified Proteins - Recombinant Protein Forum. Discuss purification, expression, and isolation of recombinant and cloned proteins here.


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Old 02-26-2008, 12:44 PM
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Post Refolding Insoluble Purified Proteins



Hello everyone,
Im trying to refold an insoluble viral glycoprotein which was expressed in E.coli, the problem is it is present in inclusion bodies.

Does anyone have experience in purifying proteins from inclusion bodies?

I want to use the protein to generate polyclonal antobodies in rabbits.

Does anyone have a protocol to refold my protein effectively and how can i check that its properly folded after refolding?

Many thanx
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Old 02-26-2008, 08:32 PM
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Default Re: Refolding Insoluble Purified Proteins

See:
"Protein expression and refolding A practical guide to getting the most out of inclusion bodies"

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Old 02-26-2008, 08:34 PM
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Default Re: Refolding Insoluble Purified Proteins

And this:
"Recombinant protein folding and misfolding in Escherichia coli"
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Old 12-22-2008, 04:48 PM
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Default Re: Refolding Insoluble Purified Proteins

Hello

If you have more than 3 cysteines, i recomend you to perform an On-column refolding, on Ion exchange or IMAC, If you use Gunidine Hydrochloryde you can use IMAC not IOn exchange, because this chaotrope is a gerd molecule, but if you use urea to dissolve your inclussion bodies you can use both IMAC or Ion Exchange.

Also you have to know that the bigger the viral protein the harder, and i assume that can aggregate because wants to form capsomers.
I see this a lot in structural "viral" proteins.
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