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Rigid, macroporous adsorbents for hydrophobic-interaction chromatography of proteins
A. E. Ivanov, L. V. Verkhovskaya, S. N. Khilko, V. P. Zubov
M. M. Shemyakin Institute of Bioorganic Chemistry, Academy of Sciences of the USSR, Moscow; Research Institute for Agricultural Biotechnology, Moscow
Abstract: Two new rigid adsorbents for hydrophobic-interaction chromatography of proteins have been synthesized. Both are based on macroporous glass with average pore diameter of 2000 Å with their surface being derivatized by chemisorbed N-butylpolyacrylamide (Butyl-PG) or N-benzylpolyacrylamide (Benzyl-PG). Studies of adsorption isoterms and of desorption conditions for model proteins (ovalbumin and γ-globuline) revealed that Butyl-PG resembles semirigid synthetic adsorbent Butyl-Toyopearl 650C (TOSOH, Japan) but has 0,6-fold capacity and 2,8-fold permeability of the latter for γ-globulin.
Several kinds of adsorbents for hydrophobic-interaction chromatography (Butyl-PG, Benzyl-PG, Butyl-Toyopearl 650C, Phenyl-Toyopearl 650S, Phenyl-Sepharose CL-4B and Octyl-Sepharose CL-4B) were compared in a model purification of high-molecular weight (M 3–4 105 Da) surface antigens of human and avian adenoviruses (hexon antigens).
All the adsorbents tested have revealed similar selectivity for the viral proteins, Butyl-PG having the maximal capacity for the hexon of egg-drop syndrom-76 virus (EDS-76). The chromatography of crude alantoic fluid from duck embryons infected with EDS-76 gave rise to 4-fold concentration of EDS-76 hexon with simultaneous 20-fold purification. The same degree of purification and 10-fold concentration were obtained during chromatography of crude cultural fluid containing hexon of human adenovirus serotype 5.
Russian Journal of Bioorganic Chemistry 1990, 16 (8):1028-1039