Ultrasensitive immuno-detection using viral nanoparticles with modular assembly using genetically-directed biotinylation

Litvinov, Julia; Hagström, Anna E. V.; Lopez, Yubitza; Adhikari, Meenu; Kourentzi, Katerina; Strych, Ulrich; Monzon, Federico A.; Foster, William; Cagle, Philip T.; Willson, Richard C.

doi:10.1007/s10529-014-1555-9

Cited by 10 publications

(12 citation statements)

References 17 publications

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“…M13 bacteriophages, like other viruses, can be chemically [22,23] or genetically [24][25][26][27] addressed and modified. M13 phages are versatile, and are useful for applications including biotemplating, phage display [28][29][30][31][32], and even tissue regeneration, as bacteriophages are innocuous to mammalian cells [33].…”

Section: Resultsmentioning

confidence: 99%

“…We produced viruses with the AviTag proteins fused to the p3 and p7 proteins (p3p7 AviTag phages), as well as phages modified either with AviTags only at p3 proteins (p3 AviTag phages) or only at the p7 proteins (p7 AviTag phages). The p3p7 AviTag phages were biotinylated ( Figure 1a) as previously reported [14,[24][25][26][27] using a modification of the method described by Litvinov et al [26]. Briefly, p3p7 AviTag phages were incubated with biotin ligase, biotin, ATP, bicine and magnesium acetate.…”

Section: Biotinylation and Characterization Of M13 Bacteriophagesmentioning

confidence: 99%

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Genetically Modified M13 Bacteriophage Nanonets for Enzyme Catalysis and Recovery

et al. 2019

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Enzyme-based biocatalysis exhibits multiple advantages over inorganic catalysts, including the biocompatibility and the unchallenged specificity of enzymes towards their substrate. The recovery and repeated use of enzymes is essential for any realistic application in biotechnology, but is not easily achieved with current strategies. For this purpose, enzymes are often immobilized on inorganic scaffolds, which could entail a reduction of the enzymes’ activity. Here, we show that immobilization to a nano-scaled biological scaffold, a nanonetwork of end-to-end cross-linked M13 bacteriophages, ensures high enzymatic activity and at the same time allows for the simple recovery of the enzymes. The bacteriophages have been genetically engineered to express AviTags at their ends, which permit biotinylation and their specific end-to-end self-assembly while allowing space on the major coat protein for enzyme coupling. We demonstrate that the phages form nanonetwork structures and that these so-called nanonets remain highly active even after re-using the nanonets multiple times in a flow-through reactor.

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Section: Resultsmentioning

confidence: 99%

Section: Biotinylation and Characterization Of M13 Bacteriophagesmentioning

confidence: 99%

Genetically Modified M13 Bacteriophage Nanonets for Enzyme Catalysis and Recovery

et al. 2019

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“…These bacteriophage (“phage”) can be genetically or chemically modified to display a wide range of functional groups for applications in bionanotechnology 45, 46 and in nanomedicine. 47 Here, phage are appealing candidates as LFA reporters because they evolved under Darwinian selection to exhibit low nonspecific binding; are readily coupled to recognition elements; 48-51 are capturable, in principle, by a single analyte and/or recognition element; 52 and are detectable at the single-reporter level. Remarkably, the viral-nanoparticle-based LFAs we have developed to date exhibit sensitivities for model analytes (viruses 41-43 and proteins 44 ) that are one-hundred-fold to one-thousand-fold greater than traditional gold-nanoparticle-based LFAs.…”

Section: Introductionmentioning

confidence: 99%

Orientational binding modes of reporters in a viral-nanoparticle lateral flow assay

Kim

Poling-Skutvik

Trabuco

et al. 2017

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Using microscopy and image analysis, we characterize binding of filamentous viral nanoparticles to a fibrous affinity matrix as models for reporter capture in a lateral flow assay (LFA). M13 bacteriophage (M13) displaying an in vivo-biotinylated peptide (AviTag) genetically fused to the M13 tail protein p3 are functionalized with fluorescent labels. We functionalize glass fiber LFA membranes with antibodies to M13, which primarily capture M13 on the major p8 coat proteins, or with avidin, which captures M13 at the biotin-functionalized tail, and compare orientational modes of reporter capture for the side- versus tip-binding recognition interactions. The number of captured M13 is greater for side-binding than for tip-binding, as expected from the number of recognition groups. Whereas two-thirds of side-bound M13 captured by an anti-M13 antibody bind immediately after colliding with the membrane, tip-bound M13 prominently exhibit three additional orientational modes that require M13 to reorient to enable binding. These results are consistent with the idea that the elongated M13 shape couples with the complex flow field in an open and disordered fibrous LFA membrane to enhance capture.

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“…To facilitate orientation‐controlled surface immobilization by the strongest noncovalent interaction between streptavidin (SAV) and biotin, the thread‐like phages have been subjected to a site‐specific biotinylation at either end of the virion . Typically, biotin accepting peptide (AP or AviTag), a well‐known 14‐mer substrate in which Lys residue can be in vivo biotinylated by bacterial biotin ligase BirA, is fused to p3 or p7.…”

Section: Introductionmentioning

confidence: 99%

Translocation Pathway‐Dependent Assembly of Streptavidin‐ and Antibody‐Binding Filamentous Virus‐Like Particles

Kim

Jeon

Hwang

et al. 2016

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Compared to well-tolerated p3 fusion, the display of fast-folding proteins fused to the minor capsid p7 and the major capsid p8, as well as in vivo biotinylation of biotin acceptor peptide (AP) fused to p7, are found to be markedly inefficient using the filamentous phage. Here, to overcome such limitations, the effect of translocation pathways, amber mutation, and phage and phagemid display systems on p7 and p8 display of antibody-binding domains are examined, while comparing the level of in vivo biotinylation of AP fused to p7 or p3. Interestingly, the in vivo biotinylation of AP occurs only in p3 fusion and the fast-folding antibody-binding scaffolds fused to p7 and p8 are best displayed via a twin-arginine translocation pathway in TG1 cells. The lower the expression level of the wild-type p8 and the smaller the size of the guest protein, the better the display of Z-domain fused to the recombinant p8. The in vivo biotinylated multifunctional filamentous virus-like particles can be vertically immobilized on streptavidin (SAV)-coated microspheres to resemble cellular microvilli-like structures, which reportedly enhance protein-protein interactions due to dramatically expanded flexible surface area.

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Ultrasensitive immuno-detection using viral nanoparticles with modular assembly using genetically-directed biotinylation

Cited by 10 publications

References 17 publications

Genetically Modified M13 Bacteriophage Nanonets for Enzyme Catalysis and Recovery

Genetically Modified M13 Bacteriophage Nanonets for Enzyme Catalysis and Recovery

Orientational binding modes of reporters in a viral-nanoparticle lateral flow assay

Translocation Pathway‐Dependent Assembly of Streptavidin‐ and Antibody‐Binding Filamentous Virus‐Like Particles

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