Fusion proteins of streptavidin and allophycocyanin alpha subunit for immunofluorescence assay

doi:10.1016/j.bej.2017.06.001

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	Fusion proteins of streptavidin and allophycocyanin alpha subunit for immunofluorescence assay
	Wu, Jing1,2,3 ; Chen, Huaxin 1,2; Zhao, Jin 1,2,3; Jiang, Peng 1,2
	2017-09-15
发表期刊	BIOCHEMICAL ENGINEERING JOURNAL
卷号	125 页码:97-103
文章类型	Article
摘要	Allophycocyanin is generally used as fluorescent label. In this study, the fusion protein (SLA) of core streptavidin from Streptomyces avidinii and allophycocyanin alpha subunit from thermophilic cyanobacterium Thermosynechococcus elongatus BP-1, together with lyase (cpcS) and phycoerythrobilin (PEB) synthesizing enzymes (Ho1 and PebS) or phycocyanobilin (PCB) synthesizing enzymes (Ho1 and PcyA), were co-expressed in Escherichia coli. The fusion proteins were purified by metal affinity chromatography. Zinc staining analysis and spectral analysis showed that the recombinant fusion proteins covalently bound PEB (denoted as SLA-PEB) or PCB (denoted as SLA-PCB). The two proteins were highly fluorescent and were able to binding biotin. a-fetoprotein (AFP), a tumor marker, was used as analyte to evaluate the application of these two fluorescent proteins in sandwich immunofluorescence assay. The measurable ranges of AFP by these proteins were 0.02-50 ng/ml with an average coefficient of variation below 15%. The detection limit for AFP detection by SLA-PEB and SLA-PCB were 0.11 and 0.35 ng/ml, respectively. The detection sensitivities were comparable to that by streptavidin-R-phycoerythrin (SA-PE). Thus, these fusion proteins would be useful fluorescent labels in immunofluorescence assay. (C) 2017 Elsevier B.V. All rights reserved.
关键词	Streptavidin Fusion Protein Biotin Immunofluorescence Assay
DOI	10.1016/j.bej.2017.06.001
收录类别	SCI
语种	英语
WOS记录号	WOS:000410017600013
引用统计	被引频次：13[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
版本	出版稿
条目标识符	http://ir.qdio.ac.cn/handle/337002/143146
专题	实验海洋生物学重点实验室
作者单位	1.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China 2.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao 266071, Peoples R China 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
第一作者单位	实验海洋生物学重点实验室
推荐引用方式 GB/T 7714	Wu, Jing,Chen, Huaxin,Zhao, Jin,et al. Fusion proteins of streptavidin and allophycocyanin alpha subunit for immunofluorescence assay[J]. BIOCHEMICAL ENGINEERING JOURNAL,2017,125:97-103.
APA	Wu, Jing,Chen, Huaxin,Zhao, Jin,&Jiang, Peng.(2017).Fusion proteins of streptavidin and allophycocyanin alpha subunit for immunofluorescence assay.BIOCHEMICAL ENGINEERING JOURNAL,125,97-103.
MLA	Wu, Jing,et al."Fusion proteins of streptavidin and allophycocyanin alpha subunit for immunofluorescence assay".BIOCHEMICAL ENGINEERING JOURNAL 125(2017):97-103.

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