Interesting Developments at the nanoparticle–protein interface: Implications for Next Generation Drug Delivery

Medintz, Igor L.

doi:10.4155/tde-2016-0035

Cited by 4 publications

(5 citation statements)

References 24 publications

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“…7 Furthermore, this work adds to the ever growing body of NP-enzyme bioconjugates and suggests the application of NP-enzyme conjugates for medical, research, industrial and commercial utility which would certainly benefit from enhanced and accelerated enzymatic activity. 25,57,58 Such enzyme-AuNP bioconjugates, and especially the phenomena that they exploit, could have a significantly broad impact on a variety of fields beyond biosensing to include environmental remediation, synthetic biology and even batch chemical processing. 59…”

Section: Discussionmentioning

confidence: 99%

Enhanced enzymatic activity from phosphotriesterase trimer gold nanoparticle bioconjugates for pesticide detection

Hondred

Breger

Garland

et al. 2017

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The rapid detection of organophosphates (OPs), a class of strong neurotoxins, is critically important for monitoring acute insecticide exposure and potential chemical warfare agent use. Herein, we improve the enzymatic activity of a phosphotriesterase trimer (PTE), an enzyme that selectively recognizes OPs directly, by conjugation with distinctly sized (i.e., 5, 10, and 20 nm diameter) gold nanoparticles (AuNPs). The number of enzymes immobilized on the AuNP was controlled by conjugating increasing molar ratios of PTE onto the AuNP surface via metal affinity coordination. This occurs between the PTE-His termini and the AuNP-displayed Ni-nitrilotriacetic acid end groups and was confirmed with gel electrophoresis. The enzymatic efficiency of the resultant PTE-AuNP bioconjugates was analyzed via enzyme progress curves acquired from two distinct assay formats that compared free unbound PTE with the following PTE-AuNP bioconjugates: (1) fixed concentration of AuNPs while increasing the bioconjugate molar ratio of PTE displayed around the AuNP and (2) fixed concentration of PTE while increasing the bioconjugate molar ratio of PTE-AuNP by decreasing the AuNP concentration. Both assay formats monitored the absorbance of p-nitrophenol that was produced as PTE hydrolyzed the substrate paraoxon, a commercial insecticide and OP nerve agent simulant. Results demonstrate a general equivalent trend between the two formats. For all experiments, a maximum enzymatic velocity (V) increased by 17-fold over free enzyme for the lowest PTE-AuNP ratio and the largest AuNP (i.e., ratio of 1 : 1, 20 nm dia. AuNP). This work provides a route to improve enzymatic OP detection strategies with enzyme-NP bioconjugates.

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Section: Discussionmentioning

confidence: 99%

Enhanced enzymatic activity from phosphotriesterase trimer gold nanoparticle bioconjugates for pesticide detection

Hondred

Breger

Garland

et al. 2017

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“…Comparison of the experimental data with the model-predicted activities at low and high enzyme concentrations supported the authors’ hypothesis that at the optimized enzyme ratios the observed improvements in catalytic activity are the product of both improved enzyme stability and substrate channeling between the QD-immobilized enzymes. Interestingly, a large number of other enzymes have shown some type of enhanced activity when assembled on both QDs and similarly sized gold nanoparticles (AuNPs), including alkaline phosphatase, phosphotriesterase and many of its engineered structural variants, luciferase, β-Gal, and acetylcholine esterase, among others. ,− ,− This phenomenon seems to arise from the unique environment around NPs, which appears to alleviate certain rate-limiting steps along with the aforementioned improvement in stability. It also certainly bodes well for their utility in any type of NP-scaffolded cascade.…”

Section: Abiotic Scaffoldsmentioning

confidence: 99%

Artificial Multienzyme Scaffolds: Pursuing in Vitro Substrate Channeling with an Overview of Current Progress

et al. 2019

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Artificial multienzyme scaffolds are being developed for in vitro cascaded biocatalytic activity and, in particular, accessing substrate channeling. This review covers progress in this field over the last ∼5 years with a specific focus on the scaffold materials themselves and the benefits they can provide for assembling multienzyme cascades in vitro. These benefits include improving biocatalytic efficiency, bypassing potential cellular toxicity, directed catalysis, modularity, incorporating enzymes from different prokaryotic and eukaryotic sources, and potentially the ability to create de novo designer cascades. We begin with an overview of the strongest impetus currently driving the rapid development of this field, namely, biomanufacturing and cell-free synthetic biology. We then discuss in detail pertinent mechanisms responsible for the benefits of artificial multienzyme scaffolds. In particular, we focus on substrate channeling, including the evolving debate about what leads to substrate channeling in artificial systemsproximity, confinement, or bothand whether sequential enzyme order is really needed. How different scaffold materials/chemistries can in turn affect enzyme activity is also discussed. The bulk of the review then details progress in the development of different biotic (e.g., cells) and abiotic (e.g., nanoparticles) scaffolding materials and is divided up by class and subtype as needed. Within each material class of scaffolds, attention is given to their inherent chemical diversity, how they are engineered, how they allow for enzymatic attachment, their ease of use, their benefits (e.g., inherent three-dimensional architecture) and liabilities where appropriate, and other relevant issues. For each scaffolding material, a detailed overview of current progress is provided using examples of multienzyme cascades and data/schematics reproduced from the literature. Special attention is also given to the use of DNA scaffolds, as they can potentially provide the most versatile designer three-dimensional scaffold architectures. Finally, a short perspective on how this rapidly moving field will evolve in the near and long terms is provided.

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“…The controlled immobilization of proteins on 2D or 3D substrates has shown great potential to revolutionize protein diagnostics and biomolecular recognition. Recent advancements in nanobiotechnology have enabled the development of strategies for the immobilization to 2D and 3D surfaces of different biological molecules such as peptides, proteins, aptamers, and nucleic acids, allowing for the fabrication of multi-functional platforms for therapeutic and diagnostic purposes, including protein microarrays [ 1 , 2 , 3 ]. Yet, the major challenges associated with protein immobilization to a solid substrate concern the preservation of protein structure-function by avoiding structural alterations of the proteins during the immobilization process as well as the steric hindrance of the recognition sites [ 4 ].…”

Section: Introductionmentioning

confidence: 99%

DNA-Directed Protein Anchoring on Oligo/Alkanethiol-Coated Gold Nanoparticles: A Versatile Platform for Biosensing Applications

et al. 2022

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Herein, we report on a smart biosensing platform that exploits gold nanoparticles (AuNPs) functionalized through ssDNA self-assembled monolayers (SAM) and the DNA-directed immobilization (DDI) of DNA-protein conjugates; a novel, high-sensitivity optical characterization technique based on a miniaturized gel electrophoresis chip integrated with online thermal lens spectrometry (MGEC-TLS), for the high-sensitivity detection of antigen binding events. Specifically, we characterized the physicochemical properties of 20 nm AuNPs covered with mixed SAMs of thiolated single-stranded DNA and bio-repellent molecules, referred to as top-terminated oligo-ethylene glycol (TOEG6), demonstrating high colloidal stability, optimal binder surface density, and proper hybridization capacity. Further, to explore the design in the frame of cancer-associated antigen detection, complementary ssDNA fragments conjugated with a nanobody, called C8, were loaded on the particles and employed to detect the presence of the HER2-ECD antigen in liquid. At variance with conventional surface plasmon resonance detection, MGEC-TLS characterization confirmed the capability of the assay to titrate the HER2-ECD antigen down to concentrations of 440 ng/mL. The high versatility of the directed protein-DNA conjugates immobilization through DNA hybridization on plasmonic scaffolds and coupled with the high sensitivity of the MGEC-TLS detection qualifies the proposed assay as a potential, easily operated biosensing strategy for the fast and label-free detection of disease-relevant antigens.

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Interesting Developments at the nanoparticle–protein interface: Implications for Next Generation Drug Delivery

Cited by 4 publications

References 24 publications

Enhanced enzymatic activity from phosphotriesterase trimer gold nanoparticle bioconjugates for pesticide detection

Enhanced enzymatic activity from phosphotriesterase trimer gold nanoparticle bioconjugates for pesticide detection

Artificial Multienzyme Scaffolds: Pursuing in Vitro Substrate Channeling with an Overview of Current Progress

DNA-Directed Protein Anchoring on Oligo/Alkanethiol-Coated Gold Nanoparticles: A Versatile Platform for Biosensing Applications

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