Quantitation of binding, recovery and desalting efficiency of peptides and proteins in solid phase extraction micropipette tips

被引:29
作者
Palmblad, M [1 ]
Vogel, JS [1 ]
机构
[1] Lawrence Livermore Natl Lab, Ctr Accelerator Mass Spectrometry, Livermore, CA 94551 USA
来源
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 2005年 / 814卷 / 02期
关键词
solid phase extraction; quantitation; recovery; peptides; proteins; accelerator mass spectrometry;
D O I
10.1016/j.jchromb.2004.10.052
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Micropipette-tip solid phase extraction (SPE) systems are common in proteomic analyses for desalting and concentrating samples for mass spectrometry, removing interferences, and increasing sensitivity. These systems are inexpensive, disposable, and highly efficient. Here, we show micropipette-tip solid phase extraction is a direct sample preparation method for (14)C-accelerator mass spectrometry (AMS), removing salts or reagent from labeled macromolecules. We compared loading, recovery and desalting efficiency in commercially available SPE microtips using (14)C-labeled peptides and proteins, AMS, and alpha spectrometry ion energy loss quantitation. The polypropylene in the tips was nearly (14)C-free and simultaneously provided low-background carrier for AMS. The silica material did not interfere with the analysis. Alpha spectrometry provided an absolute measurement of desalting efficiency. (C) 2004 Elsevier B.V. All rights reserved.
引用
收藏
页码:309 / 313
页数:5
相关论文
共 17 条
[2]   RADIOLABELING OF PROTEINS BY REDUCTIVE ALKYLATION WITH (C-14) FORMALDEHYDE AND SODIUM CYANOBOROHYDRIDE [J].
DOTTAVIOMARTIN, D ;
RAVEL, JM .
ANALYTICAL BIOCHEMISTRY, 1978, 87 (02) :562-565
[3]   Examination of micro-tip reversed-phase liquid chromatographic extraction of peptide pools for mass spectrometric analysis [J].
Erdjument-Bromage, H ;
Lui, M ;
Lacomis, L ;
Grewal, A ;
Annan, RS ;
McNulty, DE ;
Carr, SA ;
Tempst, P .
JOURNAL OF CHROMATOGRAPHY A, 1998, 826 (02) :167-181
[4]  
Gobom J, 1999, J MASS SPECTROM, V34, P105, DOI 10.1002/(SICI)1096-9888(199902)34:2<105::AID-JMS768>3.0.CO
[5]  
2-4
[6]   α-particle energy loss measurement of microgram depositions of biomolecules [J].
Grant, PG ;
Palmblad, M ;
Murov, S ;
Hillegonds, DJ ;
Ueda, DL ;
Vogel, JS ;
Bench, G .
ANALYTICAL CHEMISTRY, 2003, 75 (17) :4519-4524
[7]   A high-throughput method for the conversion of CO2 obtained from biochemical samples to graphite in septa-sealed vials for quantification of 14C via accelerator mass spectrometry [J].
Ognibene, TJ ;
Bench, G ;
Vogel, JS ;
Peaslee, GF ;
Murov, S .
ANALYTICAL CHEMISTRY, 2003, 75 (09) :2192-2196
[8]   A new accelerator mass spectrometry system for 14C-quantification of biochemical samples [J].
Ognibene, TJ ;
Bench, G ;
Brown, TA ;
Peaslee, GF ;
Vogel, JS .
INTERNATIONAL JOURNAL OF MASS SPECTROMETRY, 2002, 218 (03) :255-264
[9]  
Palmblad M, 2000, RAPID COMMUN MASS SP, V14, P1029, DOI 10.1002/1097-0231(20000630)14:12<1029::AID-RCM984>3.0.CO
[10]  
2-#