Retention behaviour of proteins under conditions of column overload in hydrophobic interaction chromatography

被引：11

作者：

Feng, WK ^{[1
]}

Zhu, X ^{[1
]}

Zhang, LH ^{[1
]}

Geng, XD ^{[1
]}

机构：

[1] NW UNIV XIAN,INST MODERN SEPARAT SCI,XIAN 710069,PEOPLES R CHINA

来源：

JOURNAL OF CHROMATOGRAPHY A | 1996年 / 729卷 / 1-2期

关键词：

retention behaviour; column overloading; preparative chromatography; proteins; cytochrome c; lysozyme;

D O I：

10.1016/0021-9673(95)01049-1

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The sample solvent has a significant effect on band profiles in preparative liquid chromatography. This effect has been described by Guiochon and Jandera in non-aqueous reversed-phase high-performance liquid chromatography. Similar behaviour was observed in hydrophobic interaction chromatography of cytochrome c and lysozyme under conditions of column overloading. If the sample solvent contains a lower concentration of salt than the starting mobile phase in gradient hydrophobic interaction chromatography, the protein peak will be broken to two peaks. One peak is longer-retarded, another is shorter-retarded by hydrophobic interaction. The latter changes its retention time with the different injection volumes. This results in a loss in the recovery of protein in preparative chromatography. The possible mechanism of the band-splitting was investigated.

引用

页码：43 / 47

页数：5

共 21 条

[1]

BARTH HG, 1986, ANAL CHEM, V58, P227

[2] CHEMICAL AND CHROMATOGRAPHIC CHARACTERIZATION OF A NEW BIOPROCESS(TM) MEDIUM FOR HYDROPHOBIC INTERACTION CHROMATOGRAPHY - BUTYL SEPHAROSE(R)-4 FAST-FLOW [J].

BERGGRUND, A ;

DREVIN, I ;

KNUUTTILA, KG ;

WARDHAMMAR, J ;

JOHANSSON, BL .

PROCESS BIOCHEMISTRY, 1994, 29 (06) :455-463

[3]

BIAN L, 1991, SEPU CHINESE J CHROM, V9, P267

[4] ADVANCED SORBENTS FOR PREPARATIVE PROTEIN SEPARATION PURPOSES [J].

BOSCHETTI, E .

JOURNAL OF CHROMATOGRAPHY A, 1994, 658 (02) :207-236

[5] STUDIES ON THE CORRELATION OF CHROMATOGRAPHIC BEHAVIOR OF BIOPOLYMERS BETWEEN REVERSED-PHASE AND HYDROPHOBIC INTERACTION CHROMATOGRAPHY - TESTING THE STOICHIOMETRIC DISPLACEMENT BETWEEN BIOPOLYMER AND DISPLACING AGENT [J].

CHANG, J ;

GUO, L ;

FENG, W ;

GENG, X .

CHROMATOGRAPHIA, 1992, 34 (11-12) :589-596

[6]

CHANG J, 1989, NW U, V9, P103

[7]

CHANG J, 1991, SEPU CHINESE J CHROM, V9, P263

[8] PREPARATIVE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY UNDER ISOCRATIC CONDITIONS .3. THE CONSEQUENCES OF 2 ADJACENT BANDS HAVING UNEQUAL COLUMN CAPACITIES [J].

COX, GB ;

SNYDER, LR .

JOURNAL OF CHROMATOGRAPHY, 1989, 483 :95-110

[9] DISPLACEMENT EFFECTS IN PREPARATIVE GRADIENT HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC SEPARATIONS [J].

COX, GB ;

SNYDER, LR .

JOURNAL OF CHROMATOGRAPHY, 1992, 590 (01) :17-33

[10]

COX GB, 1989, J CHROMATOGR, V484, P95

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