Ready-made matrix-assisted laser desorption/ionization target plates coated with thin matrix layer for automated sample deposition in high-density array format

被引:41
作者
Miliotis, T
Kjellström, S
Nilsson, J
Laurell, T
Edholm, LE
Marko-Varga, G [1 ]
机构
[1] AstraZeneca R&D Lund, Mol Sci, SE-22187 Lund, Sweden
[2] Lund Univ, Dept Analyt Chem, SE-22100 Lund, Sweden
[3] Lund Inst Technol, Dept Elect Measurements, SE-22100 Lund, Sweden
关键词
D O I
10.1002/rcm.542
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The methodology for ready-made matrix-assisted laser desorption/ionization (MALDI) target plates covered with an optimized thin layer consisting of matrix and nitrocellulose has been developed. Piezoelectric microdispensing enabled sample depositions in a high-density array format of 2000 sample depositions on a conventionally sized target plate (45 x 47 mm.). The sample depositions were made reproducibly in a fully automated mode by using an in-house developed computer-controlled piezoelectric flow-through microdispenser. Additionally, the piezoelectric technique facilitated significant analyte enrichment that increased the detection sensitivity. The MS signal was obtained rapidly, generally within ten laser pulses. An airbrush device was used to generate a fine spray of matrix and nitrocellulose dissolved in acetone. The acetone evaporated instantly when reaching the target plate leaving the entire surface with a thin and uniform matrix/nitrocellulose coating consisting of very small crystals of matrix embedded in the nitrocellulose. These crystals acted as a seed-layer on subsequent analyte depositions, rendering homogeneous sample spots when using alpha-cyano-4-hydroxycinnamic acid (CHCA) as matrix. The relative standard deviation of the signal intensity between spots was (20-30)% (n = 30). The detection sensitivity was improved by restricting the sample spot diameter to 300 mum. The spot size was affected by the deposition rate and the evaporation rate of the dispensed sample volume. Mass spectra of a 25-amol peptide mixture deposition were successfully recorded. Copyright (C) 2001 John Wiley Sons, Ltd.
引用
收藏
页码:117 / 126
页数:10
相关论文
共 29 条
[11]  
Lake DA, 2000, RAPID COMMUN MASS SP, V14, P1008, DOI 10.1002/(SICI)1097-0231(20000615)14:11<1008::AID-RCM979>3.0.CO
[12]  
2-T
[13]   Design and development of a silicon microfabricated flow-through dispenser for on-line picolitre sample handling [J].
Laurell, T ;
Wallman, L ;
Nilsson, J .
JOURNAL OF MICROMECHANICS AND MICROENGINEERING, 1999, 9 (04) :369-376
[14]   MALDI on a chip: Analysis of arrays of low femtomole to subfemtomole quantities of synthetic oligonucleotides and DNA diagnostic products dispensed by a piezoelectric pipet [J].
Little, DP ;
Cornish, TJ ;
ODonnell, MJ ;
Braun, A ;
Cotter, RJ ;
Koster, H .
ANALYTICAL CHEMISTRY, 1997, 69 (22) :4540-4546
[15]   USE OF MASS-SPECTROMETRIC MOLECULAR-WEIGHT INFORMATION TO IDENTIFY PROTEINS IN SEQUENCE DATABASES [J].
MANN, M ;
HOJRUP, P ;
ROEPSTORFF, P .
BIOLOGICAL MASS SPECTROMETRY, 1993, 22 (06) :338-345
[16]   Protein identification platform utilizing micro dispensing technology interfaced to matrix-assisted laser desorption ionization time-of-flight mass spectrometry [J].
Miliotis, T ;
Kjellström, S ;
Önnerfjord, P ;
Nilsson, J ;
Laurell, T ;
Edholm, LE ;
Marko-Varga, G .
JOURNAL OF CHROMATOGRAPHY A, 2000, 886 (1-2) :99-110
[17]  
Miliotis T, 2000, J MASS SPECTROM, V35, P369, DOI 10.1002/(SICI)1096-9888(200003)35:3<369::AID-JMS944>3.0.CO
[18]  
2-N
[19]   SAMPLE IMMOBILIZATION PROTOCOLS FOR MATRIX-ASSISTED LASER-DESORPTION MASS-SPECTROMETRY [J].
MOCK, KK ;
SUTTON, CW ;
COTTRELL, JS .
RAPID COMMUNICATIONS IN MASS SPECTROMETRY, 1992, 6 (04) :233-238
[20]  
Önnerfjord P, 1999, RAPID COMMUN MASS SP, V13, P315