Process for Purification of Human Growth Hormone
OwnerDipanwita Maiti, Shrikant Mishra +4inventors
InventorsDipanwita Maiti, Shrikant Mishra, Laxmi Srinivas Rao, Milind Prabhakar Niphadkar, Ahmed Monsur Borbhuiya, Madhava Yada Rao
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Abstract
Adding enough organic solvent to hydrophobic interactive chromatography elution buffer eliminates the need to also add detergent to the buffer when separating poly-peptides. For example, adding about 50% acetonitrile to detergent-free elution buffer enaibls one to separate full-length human growth hormone from its various truncated forms, to obtain hGH with a purity of >99.5%. This technique is useful to purify polypeptide where detergent to contamination in the resulting polypeptide is undesirable.
Claims19
1-11. (canceled)
A hydrophobic interaction chromatography process comprising:
- (i) obtaining a hydrophobic interaction chromatography column;
- (ii) loading onto said hydrophobic interaction chromatography column a sample comprising a first polypeptide and a second polypeptide;
- and
- (iii) eluting at least one of said first polypeptide and said second polypeptide from said hydrophobic interaction chromatography column with an elution buffer,
- (a) said elution buffer being substantially free of detergent,
- (b) said elution buffer comprised of at least about 10% v/v. concentration of organic solvent.
The process of claim 1, wherein said organic solvent is selected from the group consisting of a C1 to C4 alcohol, and acetonitie.
The process of claim 23, wherein said attached hydrophobic ligand comprises phenyl.
In a process for purifying a polypeptide by using hydrophobic interaction chromatography, the improvement comprising:
- using an elution buffer which is both substantially free of detergent, and which includes at least about 10% v/v concentration of organic solvent.
Dependencies are inferred from the claim text.