Characterisation of cholera toxin by liquid chromatography - Electrospray mass spectrometry

被引:16
作者
van Baar, BLM [1 ]
Hulst, AG [1 ]
Wils, ERJ [1 ]
机构
[1] TNO, Prins Maurits Lab, NL-2280 AA Rijswijk, Netherlands
关键词
D O I
10.1016/S0041-0101(98)00136-6
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
Cholera toxin, one of the toxins that may be generated by various strains of the bacterium Vibrio cholerae, can be considered as a substance possibly used in biological warfare. The possibilities of characterising the toxin by liquid chromatography electrospray mass spectrometry (LC-ES-MS) were investigated. The toxin can be detected by flow-injection (FIA) ES-MS of a dialysed solution and observation of the charge envelope signals of its A-unit and B-chain protein; sufficient information for identification by the molecular mass of either protein could be obtained for quantities in the order of 10 fmol. Confirmatory analysis was carried out by 2-mercaptoethanol reduction and FIA-ES-MS detection of the product proteins or by tryptic digest LC-ES-MS with ion chromatogram detection of most of the tryptic fragments of the A-unit and B-chain from the singly, doubly or triply charged ion signals. The confirmatory tryptic digest LC-ES-MS analysis could be achieved with quantities as low as 1 pmol. Possible biovariations in the toxin can mostly be determined by sequencing, where the amino acid composition of tryptic fragments of the A1-chain, T5 and T15, and of the B-chain, T1, T4 and T5, cover all known biovariations. Partial sequencing of cholera toxin, originating from a classical strain, O1/569B, was achieved by LC-ES-MS/MS of most tryptic fragments larger than three amino acid residues. (C) 1998 Elsevier Science Ltd. All rights reserved.
引用
收藏
页码:85 / 108
页数:24
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