Peptide quantification by matrix-assisted laser desorption ionisation time-of-flight mass spectrometry: Investigations of the cyclotide kalata B1 in biological fluids

被引:24
作者
Colgrave, ML [1 ]
Jones, A [1 ]
Craik, DJ [1 ]
机构
[1] Univ Queensland, Inst Mol Biosci, ARC Special Res Ctr Funct & Appl Genom, Brisbane, Qld 4072, Australia
关键词
quantification; cyclotides; kalata B1; circular proteins; LC/MS; MALDI-TOF mass spectrometry;
D O I
10.1016/j.chroma.2005.07.094
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A rapid method has been developed for the quantification of the prototypic cyclotide kalata B I in water and plasma utilizing matrix-assisted laser desorption ionisation time-of-flight (MALDI-TOF) mass spectrometry. The unusual structure of the cyclotides means that they do not ionise as readily as linear peptides and as a result of their low ionisation efficiency, traditional LC/MS analyses were not able to reach the levels of detection required for the quantification of cyclotides in plasma for pharmacokinetic studies. MALDI-TOF-MS analysis showed linearity (R-2 > 0.99) in the concentration range 0.05-10 mu g/mL with a limit of detection of 0.05 mu g/mL (9 fmol) in plasma. This paper highlights the applicability of MALDI-TOF mass spectrometry for the rapid and sensitive quantification of peptides in biological samples without the need for extensive extraction procedures. (c) 2005 Elsevier B.V. All rights reserved.
引用
收藏
页码:187 / 193
页数:7
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