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Micellar electrokinetic chromatography and laser induced fluorescence detection of botulinum neurotoxin type a activity using a dual-labelled substrate

文献类型:期刊论文

作者Wei, Hongping1; Kang, Yan2; Zhang, Zhi-ping1; Cui, Zong-qiang1; Zhou, Yafeng1; Zhang, Xian-En1
刊名International journal of environmental analytical chemistry
出版日期2008
卷号88期号:13页码:947-956
关键词Botulinum neurotoxin a activity Mekc Lif Dual labelling
ISSN号0306-7319
DOI10.1080/03067310802065208
通讯作者Zhang, xian-en(zhangxe@most.cn)
英文摘要The activity of botulinum neurotoxin type a (bont a) can be measured by monitoring the toxin's endopeptidase reaction with its peptide substrate. in this report, a sensitive and simple capillary electrophoresis (ce) method for analysing bont a activity was developed using a peptide substrate labelled with fluorescein isothiocynate (fitc) at the n-terminal and biotin at the c-terminal. this dual labelling enables not only highly sensitive laser induced fluorescence (lif) detection of the reaction product, but also good analytical separation of the product from the peptide substrate by micellar electrokinetic chromatography (mekc). the separation between the product peak and the substrate peak was approximately 5 min using the dual-labelled substrate, while just about 1 min using the fitc-labelled substrate without biotinylation. using the current assay method, bont a with concentration as low as 0.1 ng ml-1 (3.6 u ml(-1) in mouse ld50) in water was detected with a s:n ratio of 3 (rsd 19%) and a linear range of four orders of magnitude. with ce's advantages of very small sample volume needed, this method may find particular applications as in assays of bont a activity in water samples and kinetic analyses of toxin activity.
WOS关键词BIOLOGICAL WEAPONS ; MASS-SPECTROMETRY ; SEROTYPE-A ; TOXIN ; ASSAYS ; DIFFERENTIATION ; SERUM
WOS研究方向Chemistry ; Environmental Sciences & Ecology
WOS类目Chemistry, Analytical ; Environmental Sciences
语种英语
WOS记录号WOS:000259649200004
出版者TAYLOR & FRANCIS LTD
URI标识http://www.irgrid.ac.cn/handle/1471x/2375460
专题武汉病毒研究所
通讯作者Zhang, Xian-En
作者单位1.Chinese Acad Sci, Wuhan Inst Virol, State Key Lab Virol, Wuhan, Peoples R China
2.Huazhong Univ Sci & Technol, Coll Life Sci & Technol, Wuhan 430074, Peoples R China
推荐引用方式
GB/T 7714
Wei, Hongping,Kang, Yan,Zhang, Zhi-ping,et al. Micellar electrokinetic chromatography and laser induced fluorescence detection of botulinum neurotoxin type a activity using a dual-labelled substrate[J]. International journal of environmental analytical chemistry,2008,88(13):947-956.
APA Wei, Hongping,Kang, Yan,Zhang, Zhi-ping,Cui, Zong-qiang,Zhou, Yafeng,&Zhang, Xian-En.(2008).Micellar electrokinetic chromatography and laser induced fluorescence detection of botulinum neurotoxin type a activity using a dual-labelled substrate.International journal of environmental analytical chemistry,88(13),947-956.
MLA Wei, Hongping,et al."Micellar electrokinetic chromatography and laser induced fluorescence detection of botulinum neurotoxin type a activity using a dual-labelled substrate".International journal of environmental analytical chemistry 88.13(2008):947-956.

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来源:武汉病毒研究所

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