Purification of insulin-like growth factor-I and related proteins using underivatized silica

• Published in: Journal of Chromatography A 1996-11, Vol.753 (1), p.73-80
• Main Authors: Reifsnyder, David H., Olson, Charles V., Etcheverry, Tina, Prashad, Hardayal, Builder, Stuart E.
• Format: Article
• Language: English
• Subjects: Analytical, structural and metabolic biochemistry; Biological and medical sciences; Chromatography, High Pressure Liquid; Ethanol - chemistry; Fundamental and applied biological sciences. Psychology; Growth factors; Humans; Insulin-Like Growth Factor Binding Protein 3 - isolation & purification; Insulin-Like Growth Factor I - isolation & purification; Protein hormones. Growth factors. Cytokines; Proteins; Recombinant Proteins - isolation & purification; Silicon Dioxide - chemistry; Sodium Chloride - chemistry; Tetramethylammonium chloride
• ISSN: 0021-9673
• DOI: 10.1016/S0021-9673(96)00549-3

Adsorption chromatography using underivatized porous glass can be an effective capture step for the purification of recombinant proteins. Classical desorption techniques using chaotropic agents or harsh chemical solvents often result in elution of inactive material and may not be economical at the process scale. More recently, elution schemes have used tetramethylammonium chloride (TMAC) to obtain biologically active material. A TMAC elution was shown to be effective in the initial purification steps for the recovery of recombinant human insulin-like growth factor-I (rhIGF-I) from an Escherichia coli fermentation broth. However, TMAC also elutes other, more hydrophobic, proteins that are difficult to remove in subsequent purification steps. This paper describes the capture of IGF-I from a crude fermentation broth and a more specific elution using a combination of ethanol and NaCl rather than TMAC. This elution also can be used with other proteins including an IGF-I binding protein (BP3) expressed in mammalian cell culture.