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Protein Refolding
Sophisticated technology, better results
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More than 50% of proteins expressed in E.Coli aggregate into insoluble inclusion bodies. Proteins form
inclusion bodies are often those that are expressed at high level. Refolding them into biologically active
conformation is an attractive solution to obtain sufficient amount of samples at low cost
EZBiolab provide refolding service to restore target protein's native conformation. Our sophisticated
approach includes applying DLS and other methods to guild refolding process.
Standard Protocol
Although there are some basic principles about protein refolding, the exact conditions for each protein can
not be predicted. Solvent conditions including salts, pH, detergents and various of additives varies from
protein to protein. Kinetic process including refolding pathway and refolding rate can also be critical.
Exhausting all combinations is neither possible nor necessary. Explore even a small portion of the possible
combination is a formidable task that will be costly. Over years, based on large number of samples have
been tested, we found a few factors that often play determining roles in refolding. Based on them, we
developed a standard refolding protocol that offer a low cost solution to protein refolding.
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For a refolding project, the target protein will first be tested according to our time tested standard protocol
that includes three refolding conditions. About 65% proteins we tested so far have been successfully
refolded under one of the conditions. The refolding is judged by protein solubility.
Custom Protocol
In cases that our standard refolding protocol fails, we provide customized protocol that includes screening
up to 18 refolding conditions.
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Successfully refolded samples (judged by solubility) will be sent to customer for functional analysis. Refolding
will be scaled up based on results feedback from the customer.
Refolding Analysis
Solubility is the most common method to determine refolding results. If a protein stays in solution, refolding
is considered being success. This simple approach sometimes is quite effective. However, it is often found
that a refolded soluble protein is still biologically inactive. This can be (1) the protein forms soluble
oligermer or (2) The protein is still in unfolding/misfolding conformation, but its concentration is too low to
above its CMC.
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If necessary, EZBiolab will apply DLS (dynamic light scattering), size exclusive chromatograph and CD to
analysis refolded protein. The results will be used to guide the further action to restore the native folding
and biological function of the target protein.
Request a quote:
1. For protein refolding project, please email us for a quote: service @ezbiolab.com
2. For protein expression and purification project contracted to us, we will send you a separate quote for
refolding with the progress report if the protein is found to be insoluble in small scale test run.
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