POLYETHYLENE MEMBRANE AS A SAMPLE SUPPORT FOR DIRECT MATRIX-ASSISTED LASER-DESORPTION IONIZATION MASS-SPECTROMETRIC ANALYSIS OF HIGH-MASS PROTEINS

被引:117
作者
BLACKLEDGE, JA [1 ]
ALEXANDER, AJ [1 ]
机构
[1] BRISTOL MYERS SQUIBB CO,ANALYT RES & DEV,PHARMACEUT RES INST,SYRACUSE,NY 13221
关键词
D O I
10.1021/ac00101a009
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) of high mass proteins (> 100 000 Da), directly deposited on polyethylene membranes, is demonstrated. The spectral quality obtained, using standard sample preparation conditions, is equal or superior to that obtained with metal sample stages. Compared to the use of poly(vinylidene difluoride) transfer membranes, this material allows the acquisition of excellent quality MALDI mass spectra from high-mass proteins with a standard UV laser. This gain in capability is not at the expense of either mass accuracy or signal reproducibility; both approach that obtained with standard sample preparations on stainless steel. In addition, for the applications shown, the use of PE as a sample support reduces the severe ion suppression effects typically observed in the MALDI analysis of high-mass mixtures. This permits more precise mass measurements to be made via the use of internal calibration and is illustrated by the mass measurement of a chimeric mouse/human antibody (MW similar to 150 000 Da) by coaddition of bovine albumin dimer (MW similar to 130 000 Da).
引用
收藏
页码:843 / 848
页数:6
相关论文
共 18 条
[1]   RAPID, SENSITIVE ANALYSIS OF PROTEIN MIXTURES BY MASS-SPECTROMETRY [J].
BEAVIS, RC ;
CHAIT, BT .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (17) :6873-6877
[2]  
BLACKLEDGE JA, IN PRESS TECHNIQUES, V6
[3]  
BODNAR WR, 1994, 42ND P ASMS C MASS S
[4]   CHARGE-STATE SHIFTING OF INDIVIDUAL MULTIPLY-CHARGED IONS OF BOVINE ALBUMIN DIMER AND MOLECULAR-WEIGHT DETERMINATION USING AN INDIVIDUAL-ION APPROACH [J].
CHENG, XH ;
BAKHTIAR, R ;
VANORDEN, S ;
SMITH, RD .
ANALYTICAL CHEMISTRY, 1994, 66 (13) :2084-2087
[5]  
CREIGHTON TE, 1984, PROTEINS STRUCTURES, P33
[6]  
GALLAGHER RT, 1991, 39TH P ASMS C MASS S, P242
[7]  
HILLENKAMP F, 1991, ANAL CHEM, V63, pA1193
[8]   PLASMA DESORPTION MASS-SPECTROMETRY OF PROTEINS TRANSFERRED FROM GELS AFTER SODIUM DODECYL-SULFATE POLYACRYLAMIDE-GEL ELECTROPHORESIS [J].
KLARSKOV, K ;
ROEPSTORFF, P .
BIOLOGICAL MASS SPECTROMETRY, 1993, 22 (08) :433-440
[9]   ELECTROSPRAY-IONIZATION MASS-SPECTROSCOPY ON HYDROPHOBIC PEPTIDES ELECTROELUTED FROM SODIUM DODECYL-SULFATE POLYACRYLAMIDE-GEL ELECTROPHORESIS APPLICATION TO THE TOPOLOGY OF THE SARCOPLASMIC-RETICULUM CA2+ ATPASE [J].
LEMAIRE, M ;
DESCHAMPS, S ;
MOLLER, JV ;
LECAER, JP ;
ROSSIER, J .
ANALYTICAL BIOCHEMISTRY, 1993, 214 (01) :50-57
[10]   PEPTIDE AND PROTEIN-ANALYSIS BY ELECTROSPRAY IONIZATION MASS-SPECTROMETRY AND CAPILLARY ELECTROPHORESIS MASS-SPECTROMETRY [J].
LOO, JA ;
UDSETH, HR ;
SMITH, RD .
ANALYTICAL BIOCHEMISTRY, 1989, 179 (02) :404-412