PROTEIN DENATURATION IN NONLINEAR ISOCRATIC AND GRADIENT ELUTION CHROMATOGRAPHY

被引:21
作者
WHITLEY, RD [1 ]
ZHANG, XM [1 ]
WANG, NHL [1 ]
机构
[1] PURDUE UNIV, SCH CHEM ENGN, W LAFAYETTE, IN 47907 USA
关键词
D O I
10.1002/aic.690400617
中图分类号
TQ [化学工业];
学科分类号
0817 ;
摘要
Protein denaturation, common in hydrophobic adsorption systems, causes misinterpretation of adsorption mechanisms, interferes with analysis in analytical chromatography, and complicates the design of large-scale adsorption processes. A detailed adsorption model isolates the effects due to denaturation from those due to mass transfer and intrinsic adsorption kinetics. The model is verified using protein gradient elution data. Simulations establish that typical symptoms of denaturation in frontal and elution chromatograms include sensitivity to changes in feed composition, column length, particle size, and operating conditions (feed size, flow rate, and column history). When a denatured species adsorbs irreversibly, the elution chromatogram shows decreasing peak area with increasing incubation time and apparent adsorption hysteresis over repeated cycles. In gradient elution, the peak elution order, resolution, and relative peak height depend highly on modulator properties and operating conditions. Interfering species limit solid-phase induced denaturation by competing for binding sites. Strategies for detecting and minimizing denaturation are proposed.
引用
收藏
页码:1067 / 1081
页数:15
相关论文
共 66 条
[51]   GRADIENT SELECTION IN REVERSED-PHASE LIQUID-CHROMATOGRAPHY [J].
SCHOENMAKERS, PJ ;
BILLIET, HAH ;
TIJSSEN, R ;
DEGALAN, L .
JOURNAL OF CHROMATOGRAPHY, 1978, 149 (FEB) :519-537
[52]  
Scopes RK., 1982, PROTEIN PURIFICATION
[53]  
Snyder L.R., 1980, HIGH PERFORMANCE LIQ, V1, P207
[54]   EFFECT OF MODULATOR SORPTION IN GRADIENT ELUTION CHROMATOGRAPHY - GRADIENT DEFORMATION [J].
VELAYUDHAN, A ;
LADISCH, MR .
CHEMICAL ENGINEERING SCIENCE, 1992, 47 (01) :233-239
[55]   ROLE OF THE MODULATOR IN GRADIENT ELUTION CHROMATOGRAPHY [J].
VELAYUDHAN, A ;
LADISCH, MR .
ANALYTICAL CHEMISTRY, 1991, 63 (18) :2028-2032
[56]  
VILLERMAUX J, 1981, PERCOLATION PROCESSE, P539
[57]   SURFACTANT-MEDIATED PROTEIN HYDROPHOBIC-INTERACTION CHROMATOGRAPHY [J].
WETLAUFER, DB ;
KOENIGBAUER, MR .
JOURNAL OF CHROMATOGRAPHY, 1986, 359 :55-60
[58]  
Whitley R.D., 1991, SEPAR TECHNOL, V1, P142
[59]   ANALYSIS OF NONEQUILIBRIUM ADSORPTION DESORPTION-KINETICS AND IMPLICATIONS FOR ANALYTICAL AND PREPARATIVE CHROMATOGRAPHY [J].
WHITLEY, RD ;
VANCOTT, KE ;
WANG, NHL .
INDUSTRIAL & ENGINEERING CHEMISTRY RESEARCH, 1993, 32 (01) :149-159
[60]   DETERMINATION OF ION-EXCHANGE EQUILIBRIUM PARAMETERS OF AMINO-ACID AND PROTEIN SYSTEMS BY AN IMPULSE-RESPONSE TECHNIQUE [J].
WHITLEY, RD ;
BROWN, JM ;
KARAJGIKAR, NP ;
WANG, NHL .
JOURNAL OF CHROMATOGRAPHY, 1989, 483 :263-287