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Fusion proteins of streptavidin and allophycocyanin alpha subunit for immunofluorescence assay
Wu, Jing1,2,3; Chen, Huaxin1,2; Zhao, Jin1,2,3; Jiang, Peng1,2
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
DOI10.1016/j.bej.2017.06.001
收录类别SCI
语种英语
WOS记录号WOS:000410017600013
引用统计
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
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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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