Abstract
Immobilized stationary phases for hydrophobic interaction chromatography (HIC) of proteins are prepared by coating macroporous silica Daisogel of different porosity with hydrophobized cellulose derivatives. The polymer adsorbed on the silica surface afterwards was cross-linked with bifunctional compounds. A uniform polymer nanocoating was indicated using the nitrogen gas adsorption method BET and scanning electron microscopy. The absence of non-specific protein sorption of the synthesized adsorbents shows that the developed polymeric coating isolates silica surface from contact with the sorbate. The retention data of bovine serum albumin (BSA) in the HIC mode on different synthesized adsorbents were evaluated. It was shown that sorption capacity of such phases may vary over a wide range and depends mainly on the substitution degree of the immobilized polymer. The dynamic sorption capacity of BSA was up to 63 mg mL−1. The results proved that the new stationary phases have significant promise for the separation and purification of proteins in the HIC mode.
Similar content being viewed by others
References
Hahn R, Deinhofer K, Machold C, Jungbauer A (2003) J Chromatogr B 790:99–114
Boschetti E (1994) J Chromatogr A 658:207–236
Petro M, Berek D (1993) Chromatographia 37:549–561
Zhou FL, Muller D, Santarelli X, Jozefonvicz J (1989) J Chromatogr 476:195–203
Brunauer S, Emmett PH, Teller E (1938) J Am Chem Soc 60:309.
Hanson M, Unger KK (1992) Trends in Anal Chem 11:368–373
Queiroz JA, Tomaz CT, Cabral JMS (2001) J Biotechnol 87:143–159
IUPAC Recommendations Pure Appl Chem (1985), 57:603
IUPAC Recommendations Pure Appl Chem (1994), 66:1739
Serys A, Liesiene J, Urbonaviciene J, Maruska A, Radzevicius K, Valatkeviciute D, Bumelis V (1994) J Liq Chromatogr Relat Technol 17:749–760
Author information
Authors and Affiliations
Corresponding author
Additional information
Presented at: 6th Balaton Symposium on High-Performance Separation Methods, Siófok, Hungary, September 7–9, 2005
Rights and permissions
About this article
Cite this article
Danilevicius, A., Niemeyer, B. & Liesiene, J. Immobilized Stationary Phases for Hydrophobic Interaction Chromatography of Proteins. Chroma 63 (Suppl 13), S75–S80 (2006). https://doi.org/10.1365/s10337-006-0715-z
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1365/s10337-006-0715-z