“Stable-on-the-Table” Biosensors: Hemoglobin-Poly  (Acrylic Acid) Nanogel BioElectrodes with High Thermal Stability and Enhanced Electroactivity

Ghimire, Ananta; Zore, Omkar V.; Thilakarathne, Vindya K.; Briand, Victoria A.; Lenehan, Patrick; Lei, Yu; Kasi, Rajeswari M.; Kumar, Challa V.

doi:10.3390/s150923868

Cited by 12 publications

(8 citation statements)

References 45 publications

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“…7B), which is a reliable measure for quantifying the intracellular metabolism in a mammalian cell line. 38,39 Optimal intracellular dehydrogenase concentration is essential for efficient metabolism in the human cells. The reduction in dehydrogenase concentration, which is a quantitative measure of reduced intracellular metabolism, confirmed the reduction in cellular activity and eventually, the cell death.…”

Section: Resultsmentioning

confidence: 99%

Nanoarmoring: strategies for preparation of multi-catalytic enzyme polymer conjugates and enhancement of high temperature biocatalysis

et al. 2017

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We report a general and modular approach for the synthesis of multi enzyme–polymer conjugates (MECs) consisting of five different enzymes of diverse isoelectric points and distinct catalytic properties conjugated within a single universal polymer scaffold. The five model enzymes chosen include glucose oxidase (GOx), acid phosphatase (AP), lactate dehydrogenase (LDH), horseradish peroxidase (HRP) and lipase (Lip). Poly(acrylic acid) (PAA) is used as the model synthetic polymer scaffold that will covalently conjugate and stabilize multiple enzymes concurrently. Parallel and sequential synthetic protocols are used to synthesise MECs, 5-P and 5-S, respectively. Also, five different single enzyme–PAA conjugates (SECs) including GOx–PAA, AP–PAA, LDH–PAA, HRP–PAA and Lip–PAA are synthesized. The composition, structure and morphology of MECs and SECs are confirmed by agarose gel electrophoresis, dynamic light scattering, circular dichroism spectroscopy and transmission electron microscopy. The bioreactor comprising MEC functions as a single biocatalyst can carry out at least five different or orthogonal catalytic reactions by virtue of the five stabilized enzymes, which has never been achieved to-date. Using activity assays relevant for each of the enzymes, for example AP, the specific activity of AP at room temperature and 7.4 pH in PB is determined and set at 100%. Interestingly, MECs 5-P and 5-S show specific activities of 1800% and 600%, respectively, compared to 100% specific activity of AP at room temperature (RT). The catalytic efficiencies of 5-P and 5-S are 1.55 × 10−3 and 1.68 × 10−3, respectively, compared to 9.11 × 10−5 for AP under similar RT conditions. Similarly, AP relevant catalytic activities of 5-P and 5-S at 65 °C show 100 and 300%, respectively, relative to native AP activity at RT as the native AP is catalytically inactive at 65 °C The catalytic activity trends suggest: (1) MECs show enhanced catalytic activities compared to native enzymes under similar assay conditions and (2) 5-S is better suited for high temperature biocatalysis, while both 5-S and 5-P are suitable for room temperature biocatalysis. Initial cytotoxicity results show that these MECs are non-lethal to human cells including human embryonic kidney [HEK] cells when treated with doses of 0.01 mg mL−1 for 72 h. This cytotoxicity data is relevant for future biological applications.

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Section: Resultsmentioning

confidence: 99%

Nanoarmoring: strategies for preparation of multi-catalytic enzyme polymer conjugates and enhancement of high temperature biocatalysis

et al. 2017

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“…In the past few decades, PE materials have emerged as a kind of soft yet functional support for not only efficient loading of enzymes but also a confined environment that typically leads to improved enzymatic activity. , Particularly, PE micro/nanogels consisting of cross-linked PE networks integrate the advanced features of both hydrogels and PEs. As hydrogels, PE nanogels show high water content, tunable chemical and physical structures, high stability, and biocompatibility. − Meanwhile, the abundant intrinsic charges of PE nanogels allow the selective and efficient loading of enzyme without disturbing their secondary structures and functions. , For example, Dubey et al constructed core–shell PNIPAm-PEI microgels, and the designed cationic PEI blocks realized an efficient loading of acetyl CoA synthetase (286 μg/mg) . Sigolaeva and co-workers prepared P(NIPAm- co -DMAEMA) microgels that allow manipulation of the location of the enzyme by tuning temperature, pH, and enzymes with different sizes. , Therefore, PE micro/nanogels demonstrate a type of advanced hydrogel particles for enzyme immobilization.…”

Section: Introductionmentioning

confidence: 99%

Regulated Polyelectrolyte Nanogels for Enzyme Encapsulation and Activation

Cai

Ding

et al. 2021

Biomacromolecules

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Polyelectrolyte (PE) nanogels consisting of crosslinked PE networks integrate the advanced features of both nanogels and PEs. The soft environment and abundant intrinsic charges are of special interest for enzyme immobilization. However, the crucial factors that regulate enzyme encapsulation and activation remain obscure to date. Herein, we synthesized cationic poly (dimethyl aminoethyl methacrylate), PDMAEMA, nanogels with well-defined size and cross-link degrees and fully investigated the effects of different control factors on lipase immobilization. We demonstrate that the cationic PDMAEMA nanogels indeed enable efficient and safe loading of anionic lipase without disturbing their structures. Strong charge interaction achieved by tuning pH and larger particle size are favorable for lipase loading, while the enhanced enzymatic activity demands nanogels with smaller size and a moderate cross-link degree. As such, PDMAEMA nanogels with a hydrodynamic radius of 35 nm and 30% cross-linker fraction display the optimal catalytic efficiency, which is fourfold of that of free lipase. Moreover, the immobilization endows enhanced enzymatic activity in a broad scope of pH, ionic strength, and temperature, demonstrating effective protection and activation of lipase by the designed nanogels. Our study validates the crucial controls of the size and structure of PE nanogels on enzyme encapsulation and activation, and the revealed findings shall be helpful for designing functional PE nanogels and boosting their applications for enzyme immobilization.

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“…Nanospheres as immunoprobes were produced using chitosan-poly(acrylic acid) nanospheres doped with copper, cadmium, lead, and zinc ions to detect electrochemical signals and react with glutaraldehyde to immobilize various tagged antibodies [ 188 ]. Hydrogen peroxide with an excellent detection limit of 0.5 μM using Met-hemoglobin was developed as an electrochemical biosensor [ 189 ].…”

Section: Biomedical Applicationsmentioning

confidence: 99%

Polyacrylic Acid Nanoplatforms: Antimicrobial, Tissue Engineering, and Cancer Theranostic Applications

et al. 2022

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Polyacrylic acid (PAA) is a non-toxic, biocompatible, and biodegradable polymer that gained lots of interest in recent years. PAA nano-derivatives can be obtained by chemical modification of carboxyl groups with superior chemical properties in comparison to unmodified PAA. For example, nano-particles produced from PAA derivatives can be used to deliver drugs due to their stability and biocompatibility. PAA and its nanoconjugates could also be regarded as stimuli-responsive platforms that make them ideal for drug delivery and antimicrobial applications. These properties make PAA a good candidate for conventional and novel drug carrier systems. Here, we started with synthesis approaches, structure characteristics, and other architectures of PAA nanoplatforms. Then, different conjugations of PAA/nanostructures and their potential in various fields of nanomedicine such as antimicrobial, anticancer, imaging, biosensor, and tissue engineering were discussed. Finally, biocompatibility and challenges of PAA nanoplatforms were highlighted. This review will provide fundamental knowledge and current information connected to the PAA nanoplatforms and their applications in biological fields for a broad audience of researchers, engineers, and newcomers. In this light, PAA nanoplatforms could have great potential for the research and development of new nano vaccines and nano drugs in the future.

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“Stable-on-the-Table” Biosensors: Hemoglobin-Poly (Acrylic Acid) Nanogel BioElectrodes with High Thermal Stability and Enhanced Electroactivity

Cited by 12 publications

References 45 publications

Nanoarmoring: strategies for preparation of multi-catalytic enzyme polymer conjugates and enhancement of high temperature biocatalysis

Nanoarmoring: strategies for preparation of multi-catalytic enzyme polymer conjugates and enhancement of high temperature biocatalysis

Regulated Polyelectrolyte Nanogels for Enzyme Encapsulation and Activation

Polyacrylic Acid Nanoplatforms: Antimicrobial, Tissue Engineering, and Cancer Theranostic Applications

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