Resolution of proteins in linear gradient elution ion-exchange and hydrophobic interaction chromatography.

The following equation was derived for the resolution, Rs, of proteins in linear gradient elution ion-exchange and hydrophobic interaction chromatography: Rs alpha [(column length.molecular diffusivity)/(slope of the gradient.column gel volume.linear mobile phase velocity.particle diameter2)]1/2. Experimental results obtained under a wide range of experimental conditions (column length, 1-30 cm; particle diameter, 37-92 micron; linear mobile phase velocity, 0.2-3.5 cm/min; temperature, 15-35 degrees C) have shown that this relationship is valid except for very short columns with a shallow slope of the gradient and for low flow-rates. The relationship is very useful for scaling-up the linear gradient elution of proteins from data obtained with small columns.

[1]  Y. Kato,et al.  Preparative high-performance ion-exchange chromatography , 1985 .

[2]  W. Pitt.  Gradient-Elution Ion Exchange Chromatography: A Digital Computer Solution of the Mathematical Model , 1976 .

[3]  P. Carroad,et al.  Estimation of diffusion coefficients of proteins , 1980 .

[4]  J. Pearson.  High-performance liquid chromatography column length designed for submicrogram scale protein isolation. , 1986, Analytical biochemistry.

[5]  Y. Kato,et al.  Packing of Toyopearl columns for gel filtration : II. Dependence of optimal packing velocity on column size , 1981 .

[6]  K. Nakanishi,et al.  Ion exchange chromatography of proteins—prediction of elution curves and operating conditions. I. Theoretical considerations , 1983, Biotechnology and bioengineering.

[7]  Shuichi Yamamoto,et al.  Purification of β-galactosidase by large-scale gradient elution ion-exchange chromatography , 1987 .

[8]  Y. Kato,et al.  Operational variables in medium-performance ion-exchange chromatography of proteins on DEAE-Toyopearl 650 , 1983 .

[9]  Kato Yoshio,et al.  Preparative high-performance hydrophobic interaction chromatography of proteins on TSKgel phenyl-5PW , 1985 .

[10]  F. Arnold,et al.  Liquid chromatography plate height equations , 1985 .

[11]  Y. Kato,et al.  Packing of Toyopearl columns for gel filtration , 1981 .

[12]  Y. Sano,et al.  Factors affecting the relationship between the plate height and the linear mobile phase velocity in gel filtration chromatography of proteins. , 1987, Journal of chromatography.

[13]  Shuichi Yamamoto,et al.  SCALING UP OF MEDIUM-PERFORMANCE GEL FILTRATION CHROMATOGRAPHY OF PROTEINS , 1986 .

[14]  F. Regnier,et al.  Variables in the high-performance anion-exchange chromatography of proteins. , 1980, Analytical biochemistry.

[15]  L. Snyder,et al.  Separation of proteins by gradient elution from ion-exchange columns : Optimizing experimental conditions , 1986 .

[16]  Differences in retention behavior between small and large molecules in ion-exchange chromatography and reversed-phase chromatography. , 1984, Analytical biochemistry.

[17]  Ryuichi Matsuno,et al.  Ion exchange chromatography of proteins—predictions of elution curves and operating conditions. II. Experimental verification , 1983, Biotechnology and bioengineering.

[18]  Y. Kato,et al.  Study of experimental conditions in high-performance ion-exchange chromatography of proteins , 1982 .