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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
Source PublicationBIOCHEMICAL ENGINEERING JOURNAL
Volume125Pages:97-103
SubtypeArticle
AbstractAllophycocyanin 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.
KeywordStreptavidin Fusion Protein Biotin Immunofluorescence Assay
DOI10.1016/j.bej.2017.06.001
Indexed BySCI
Language英语
WOS IDWOS:000410017600013
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Cited Times:3[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Version出版稿
Identifierhttp://ir.qdio.ac.cn/handle/337002/143146
Collection实验海洋生物学重点实验室
Affiliation1.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
First Author Affilication中国科学院海洋研究所
Recommended Citation
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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