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
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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 |
DOI | 10.1080/03067310802065208 |
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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