PURIFICATION OF OVALBUMIN AND LYSOZYME FROM A COMMERCIAL PRODUCT BY RECYCLING ISOTACHOPHORESIS

被引:17
作者
CASLAVSKA, J [1 ]
GEBAUER, P [1 ]
THORMANN, W [1 ]
机构
[1] UNIV BERN,DEPT CLIN PHARMACOL,MURTENSTR 35,CH-3010 BERN,SWITZERLAND
来源
JOURNAL OF CHROMATOGRAPHY | 1991年 / 585卷 / 01期
关键词
D O I
10.1016/0021-9673(91)85068-Q
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
The aim of this work was to test the suitability of using recycling isotachophoresis (RITP) for the purification of ovalbumin (OVA) and/or lysozyme (LYSO) from a commercial OVA product containing LYSO and conalbumin (CAL) as major proteinaceous impurities. The search for suitable electrolyte systems and spacers was carried out by capillary isotachophoresis. RITP was performed in a recycling free-flow focusing apparatus in the batch mode with immobilization of the advancing zone structure via a controlled counter-flow. Typically 700 mg of the commercial product were processed within 2 h. Enhancement of the sample load was achieved by a feed of sample under counterflow control. The collected fractions were analysed separately for conductivity, pH and ultraviolet absorption, and selected fractions were characterized by analytical capillary electrophoretic methods. All three proteins could be separated and fractionated using suitable spacers. Depending on the chosen conditions either OVA or LYSO could be purified in amounts larger than milligrams per hour (OVA 300 mg/h; LYSO 10 mg/h). The instability of CAL in solution prevented its isolation in the investigated configurations.
引用
收藏
页码:145 / 152
页数:8
相关论文
共 12 条
  • [1] BIER M, 1979, ELECTROFOCUS 78, P35
  • [2] RECYCLING AND SCREEN-SEGMENTED COLUMN ISOTACHOPHORESIS, 2 FREE-FLUID APPROACHES FOR FRACTIONATION OF PROTEINS
    CASLAVSKA, J
    GEBAUER, P
    ODERMATT, A
    THORMANN, W
    [J]. JOURNAL OF CHROMATOGRAPHY, 1991, 545 (02): : 315 - 329
  • [3] EGG PROTEINS
    FEVOLD, HL
    [J]. ADVANCES IN PROTEIN CHEMISTRY, 1951, 6 : 187 - 252
  • [4] ISOTACHOPHORESIS OF PROTEINS IN UNCOATED OPEN-TUBULAR FUSED-SILICA CAPILLARIES WITH A SIMPLE APPROACH FOR COLUMN CONDITIONING
    GEBAUER, P
    THORMANN, W
    [J]. JOURNAL OF CHROMATOGRAPHY, 1991, 558 (02): : 423 - 429
  • [5] ISOTACHOPHORESIS IN OPEN-TUBULAR FUSED-SILICA CAPILLARIES WITH ON-COLUMN MULTIWAVELENGTH DETECTION
    GEBAUER, P
    THORMANN, W
    [J]. JOURNAL OF CHROMATOGRAPHY, 1991, 545 (02): : 299 - 305
  • [6] FREE-FLOW ELECTROPHORESIS FOR THE PURIFICATION OF PROTEINS .1. ZONE ELECTROPHORESIS AND ISOTACHOPHORESIS
    HOFFSTETTERKUHN, S
    KUHN, R
    WAGNER, H
    [J]. ELECTROPHORESIS, 1990, 11 (04) : 304 - 309
  • [7] HOLLOWAY CJ, 1984, METHOD ENZYMOL, V104, P281
  • [8] FREE-FLOW ELECTROPHORESIS AS A METHOD FOR THE PURIFICATION OF ENZYMES FROM ESCHERICHIA-COLI CELL EXTRACT
    KUHN, R
    WAGNER, H
    [J]. ELECTROPHORESIS, 1989, 10 (03) : 165 - 172
  • [9] NOWICKA G, 1990, J LIPID RES, V31, P1173
  • [10] PURIFICATION OF MONOCLONAL-ANTIBODIES AGAINST THE LOW-DENSITY LIPOPROTEIN RECEPTOR BY PREPARATIVE ISOTACHOPHORESIS
    SCHMITZ, G
    BOTTCHER, A
    KAHL, HG
    BRUNING, T
    [J]. JOURNAL OF CHROMATOGRAPHY-BIOMEDICAL APPLICATIONS, 1988, 431 (02): : 327 - 342