Micropreparation of peptides by capillary electrophoresis for matrix assisted laser desorption mass spectrometry

被引:9
作者
Bergman, AC [1 ]
Bergman, T [1 ]
机构
[1] KAROLINSKA INST,DEPT MED BIOCHEM & BIOPHYS,S-17177 STOCKHOLM,SWEDEN
关键词
capillary electrophoresis; matrix assisted laser desorption mass spectrometry; peptide; micropreparation; hydrochloric acid; trifluoroacetic acid;
D O I
10.1016/S0014-5793(96)01083-6
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
In the separation of peptides by capillary electrophoresis and analysis by matrix assisted laser desorption mass spectrometry, strong suppression of the mass spectrometric signals is a problem with many common electrolytes used in the separation step such as sodium phosphate. We describe an approach employing individual electrolytes selected for highest performance in each process. Suppression with samples collected into phosphate buffers is avoided when citrate, trifluoroacetic acid or hydrochloric acid is used for collection, while phosphate still provides excellent resolution in the capillary. Low concentrations of hydrochloric acid added to the sample/matrix mixture generate essentially adduct-free mass spectra with better signal-to-noise ratios and detection limits (fmol range) than those obtained with citrate or trifluoroacetic acid. Addition of 0.25% ethylene glycol to both the phosphate electrolyte and the sample improves peak shape and resolution, and is crucial for preparative separations in large diameter capillaries.
引用
收藏
页码:45 / 49
页数:5
相关论文
共 23 条
  • [1] THE USE OF CAPILLARY ELECTROPHORESIS IN A MICROPREPARATIVE MODE - METHODS AND APPLICATIONS
    ALBIN, M
    CHEN, SM
    LOUIE, A
    PAIRAUD, C
    COLBURN, J
    WIKTOROWICZ, J
    [J]. ANALYTICAL BIOCHEMISTRY, 1992, 206 (02) : 382 - 388
  • [2] DIRECT ANALYSIS OF PEPTIDES AND AMINO-ACIDS FROM CAPILLARY ELECTROPHORESIS
    BERGMAN, T
    AGERBERTH, B
    JORNVALL, H
    [J]. FEBS LETTERS, 1991, 283 (01) : 100 - 103
  • [3] BERGMAN T, 1993, METHODS IN PROTEIN SEQUENCE ANALYSIS, P21
  • [4] A SYNTHETIC PEPTIDE ENCOMPASSING THE BINDING-SITE OF THE 2ND ZINC ATOM (THE STRUCTURAL ZINC) OF ALCOHOL-DEHYDROGENASE
    BERGMAN, T
    JORNVALL, H
    HOLMQUIST, B
    VALLEE, BL
    [J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 1992, 205 (02): : 467 - 470
  • [5] MULTIPLE SEQUENTIAL FRACTION COLLECTION OF PEPTIDES AND GLYCOPEPTIDES BY HIGH-PERFORMANCE CAPILLARY ELECTROPHORESIS
    BOSS, HJ
    ROHDE, MF
    RUSH, RS
    [J]. ANALYTICAL BIOCHEMISTRY, 1995, 230 (01) : 123 - 129
  • [6] CHENG YF, 1993, BIOTECHNIQUES, V14, P51
  • [7] CORRADINI D, 1996, LC GC EUR, V9, P34
  • [8] PREPARATIVE CAPILLARY ELECTROPHORESIS BASED ON ADSORPTION OF THE SOLUTES (PROTEINS) ONTO A MOVING BLOTTING MEMBRANE AS THEY MIGRATE OUT OF THE CAPILLARY
    ERIKSSON, KO
    PALM, A
    HJERTEN, S
    [J]. ANALYTICAL BIOCHEMISTRY, 1992, 201 (02) : 211 - 215
  • [9] PROTOCOL FOR RESOLVING PROTEIN MIXTURES IN CAPILLARY ZONE ELECTROPHORESIS
    GORDON, MJ
    LEE, KJ
    ARIAS, AA
    ZARE, RN
    [J]. ANALYTICAL CHEMISTRY, 1991, 63 (01) : 69 - 72
  • [10] Hathaway Gary M., 1992, Methods (Orlando), V4, P244, DOI 10.1016/1046-2023(92)90039-B