Aptamer-Phage Reporters for Ultrasensitive Lateral Flow Assays

Long fascinating to biologists, viruses offer nanometer-scale benchtops for building molecular-scale devices and materials. Viruses tolerate a wide-range of chemical modifications including reaction conditions, pH values, and temperatures. Recent examples of non-genetic manipulation of viral surfaces have extended viruses into applications ranging from biomedical imaging, drug delivery, tissue regeneration, and biosensors to materials for catalysis and energy generation. Chemical reactions on the phage surface include both covalent and non-covalent modifications, including some applied in conjunction with genetic modifications. Here, we survey viruses chemically augmented with capabilities limited only by imagination.

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“…58 In a subsequent study, the detection antibody was replaced with an aptamer linked to P3 through a biotinylated PEG linker and neutravidin. 59 …”

Section: Applications Of Modified Phagementioning

confidence: 99%

Chemically Modifying Viruses for Diverse Applications

Mohan

Weiss

2016

ACS Chem. Biol.

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“…27-29 M13 phage are filamentous viral nanoparticles (900 nm long and 6 nm diameter) that infect and replicate within Escherichia coli . Phage produced within E. coli are very uniform in size and shape, and can readily serve as a scaffold on which to attach multiple biorecognition and read-out-signal molecules.…”

Section: Introductionmentioning

confidence: 99%

Increasing Binding Efficiency via Reporter Shape and Flux in a Viral Nanoparticle Lateral-Flow Assay

Kim

Kourentzi

et al. 2017

ACS Appl. Mater. Interfaces

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To identify factors controlling the performance of reporter particles in a sensitive lateral-flow assay (LFA), we investigated the effect of the flux and shape of filamentous bacteriophage (phage) on the performance of phage LFAs. Phage of three different lengths and diameters were modified with biotin and AlexaFluor 555 as binding and read-out elements, respectively. The binding efficiencies of the functionalized phage were tested in a fibrous glass LFA membrane modified with avidin. The total binding rate, quantified using real-time particle counting and particle image velocimetry, decreased monotonically with the average bulk flux of phage through the membrane. At the pore scale more phage bound in regions with faster local flow, confirming that both average and local flux increased binding. The number of bound phage increased with the aspect ratio of the phage and scaled with the phage surface area, consistent with a binding interaction controlled by the number of recognition elements on the surface. Together, these results indicate that increasing the likelihood that recognition elements on the surface of phage encounter the fibers enhances the assay binding efficiency and suggests one origin for the improved performance of nonspherical phage reporters.

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“…Recently, we have employed M13 bacteriophage (M13) decorated with enzymes as viral-nanoparticle reporters in ultrasensitive LFAs. 41-44 Filamentous M13 is highly anisotropic, with length of approximately 900 nm (depending on genome size) and width of approximately 6.6 nm. 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.…”

Section: Introductionmentioning

confidence: 99%

“…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. The mechanisms underlying the ultrasensitivity of viral-nanoparticle-based LFAs, however, remain poorly understood.…”

Section: Introductionmentioning

confidence: 99%

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

Kim

Poling-Skutvik

Trabuco

et al. 2017

Analyst

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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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Aptamer-Phage Reporters for Ultrasensitive Lateral Flow Assays

Cited by 35 publications

References 38 publications

Chemically Modifying Viruses for Diverse Applications

Chemically Modifying Viruses for Diverse Applications

Increasing Binding Efficiency via Reporter Shape and Flux in a Viral Nanoparticle Lateral-Flow Assay

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

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