what is the most suitable purification method for the commercial production of bioactive peptide?
Dear Yongjin
Do you know the molecular size and and amino acid sequence/ratios of the peptide?
If the peptide is small and does not carry too many modifications it could be separated by reverse phase chromatography without destroying the activity. Care should be taken if there are cysteine residues as these can cross-link with other peptides and the buffer used can play a role in the success of the separation.
The alternative would be size exclusion or gel filtration chromatography or the use of a unique property of the peptide that can be used to capture the peptide selectively onto a gel. There are many different characteristics that would be suitable for this type of selective capture onto a soft gel column which is well suited for commercial scale isolations and purification of peptides and proteins.
You will need to make choices based on the scale of the operation, the purity of the feed stock and degree of purification required, what the peptide will be used for.
More information could help to give a more selective answer.
Dear Yongjin
Do you know the molecular size and and amino acid sequence/ratios of the peptide?
If the peptide is small and does not carry too many modifications it could be separated by reverse phase chromatography without destroying the activity. Care should be taken if there are cysteine residues as these can cross-link with other peptides and the buffer used can play a role in the success of the separation.
The alternative would be size exclusion or gel filtration chromatography or the use of a unique property of the peptide that can be used to capture the peptide selectively onto a gel. There are many different characteristics that would be suitable for this type of selective capture onto a soft gel column which is well suited for commercial scale isolations and purification of peptides and proteins.
You will need to make choices based on the scale of the operation, the purity of the feed stock and degree of purification required, what the peptide will be used for.
More information could help to give a more selective answer.
As Duncan wrote, more details would be useful (pI, Mw, any tags?). Otherwise it will be trial and error.
But some non-specific chromatography methods (ionex, hydrophobic, size exclusion) will help in any case, only their rpofitability will vary.
We solved the structure of some bioactive peptides originally isolated by the group of Prof. Viktor Mutt at the Karolinska Institute. Prof. Mutt's laboratory was unique in the ability to perform process scale purification of proteins and peptides from natural sources. Being used to semi-preparative and analytical scale methods, visiting Prof. Mutts warehouse-like facility with large vats and meter-wide columns was an experience. While the purification method depends ultimately on the characteristics of the peptide, common to his approach to early stages of purification was chemical extraction, concentration, and ion exchange chromatography. Extraction and ion exchange are economical and scale well. Below is an example reference that may help you. Good Luck.
Eur. J. Biochem. 211,377-380 (1993)
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