Abstract
In order to obtain an ultra-sensitive molecular biosensor, we designed an auto-biotinylated bifunctional protein nanowire (bFPNw) based on the self-assembly of a yeast amyloid protein, Sup35, to which protein G and a biotin acceptor peptide (BAP) were genetically fused. These auto-biotinylated bFPNws can transfer hundreds of commercially available diagnostic enzymes to an antigen-antibody complex via the biotin-avidin system, greatly enhancing the sensitivity of immune-biosensing. Compared to our previously reported seeding-induced bFPNws (Men et al., 2009), these auto-biotinylated bFPNws gave greater signal amplification, reduced non-specific binding and improved stability. The auto-biotinylated self-assembled bFPNw molecular biosensors were applied to detect Yersinia pestis (Y. pestis) F1 antigen and showed a 2000- to 4000-fold increase in sensitivity compared to traditional immunoassays, demonstrating the potential use of these self-assembling protein nanowires in biosensing.
Copyright © 2010 Elsevier B.V. All rights reserved.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Antigens, Bacterial / analysis
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Awards and Prizes
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Bacterial Proteins / analysis
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Bacterial Proteins / chemistry
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Biosensing Techniques / instrumentation*
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Biosensing Techniques / methods
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Biosensing Techniques / statistics & numerical data
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Biotinylation
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Cross-Linking Reagents / chemistry
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Immunoassay / instrumentation*
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Immunoassay / methods
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Immunoassay / statistics & numerical data
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Nanowires / chemistry*
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Nanowires / ultrastructure
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Peptide Termination Factors / chemistry
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Peptide Termination Factors / ultrastructure
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Protein Multimerization
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / ultrastructure
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Saccharomyces cerevisiae Proteins / chemistry
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Saccharomyces cerevisiae Proteins / ultrastructure
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Yersinia pestis / immunology
Substances
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Antigens, Bacterial
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Bacterial Proteins
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Cross-Linking Reagents
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IgG Fc-binding protein, Streptococcus
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Peptide Termination Factors
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Recombinant Fusion Proteins
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SUP35 protein, S cerevisiae
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Saccharomyces cerevisiae Proteins
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caf1 protein, Yersinia pestis