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1.
Membranes (Basel) ; 11(10)2021 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-34677509

RESUMO

Membrane proteins are involved in many aspects of cellular biology; for example, they regulate how cells interact with their environment, so such proteins are important drug targets. The rapid advancement in the field of immune effector cell therapy has been expanding the horizons of synthetic membrane receptors in the areas of cell-based immunotherapy and cellular medicine. However, the investigation of membrane proteins, which are key constituents of cells, is hampered by the difficulty and complexity of their in vitro synthesis, which is of unpredictable yield. Cell-free synthesis is herein employed to unravel the impact of the expression construct on gene transcription and translation, without the complex regulatory mechanisms of cellular systems. Through the systematic design of plasmids in the immediacy of the start of the target gene, it was possible to identify translation initiation and the conformation of mRNA as the main factors governing the cell-free expression efficiency of the human voltage-dependent anion channel (VDAC), which is a relevant membrane protein in drug-based therapy. A simple translation initiation model was developed to quantitatively assess the expression potential for the designed constructs. A scoring function that quantifies the feasibility of the formation of the translation initiation complex through the ribosome-mRNA hybridization energy and the accessibility of the mRNA segment binding to the ribosome is proposed. The scoring function enables one to optimize plasmid sequences and semi-quantitatively predict protein expression efficiencies. This scoring function is publicly available as webservice XenoExpressO at University of Vienna, Austria.

2.
Front Mol Biosci ; 8: 677547, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34631792

RESUMO

Several attempts have been made to encapsulate indomethacin (IND), to control its sustained release and reduce its side effects. To develop a successful formulation, drug release from a polymeric matrix and subsequent biodegradation need to be achieved. In this study, we focus on combining microfluidic and artificial intelligence (AI) technologies, alongside using biomaterials, to generate drug-loaded polymeric microparticles (MPs). Our strategy is based on using Poly (D,L-lactide-co-glycolide) (PLGA) as a biodegradable polymer for the generation of a controlled drug delivery vehicle, with IND as an example of a poorly soluble drug, a 3D flow focusing microfluidic chip as a simple device synthesis particle, and machine learning using artificial neural networks (ANNs) as an in silico tool to generate and predict size-tunable PLGA MPs. The influence of different polymer concentrations and the flow rates of dispersed and continuous phases on PLGA droplet size prediction in a microfluidic platform were assessed. Subsequently, the developed ANN model was utilized as a quick guide to generate PLGA MPs at a desired size. After conditions optimization, IND-loaded PLGA MPs were produced, and showed larger droplet sizes than blank MPs. Further, the proposed microfluidic system is capable of producing monodisperse particles with a well-controllable shape and size. IND-loaded-PLGA MPs exhibited acceptable drug loading and encapsulation efficiency (7.79 and 62.35%, respectively) and showed sustained release, reaching approximately 80% within 9 days. Hence, combining modern technologies of machine learning and microfluidics with biomaterials can be applied to many pharmaceutical applications, as a quick, low cost, and reproducible strategy.

3.
Front Mol Biosci ; 8: 651232, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33869287

RESUMO

We report on the fabrication and characterisation of graphene field-effect transistor (GFET) biosensors for the detection of Clusterin, a prominent protein biomarker of Alzheimer's disease (AD). The GFET sensors were fabricated on Si/SiO2 substrate using photolithographic patterning and metal lift-off techniques with evaporated chromium and sputtered gold contacts. Raman Spectroscopy was performed on the devices to determine the quality of the graphene. The GFETs were annealed to improve their performance before the channels were functionalized by immobilising the graphene surface with linker molecules and anti-Clusterin antibodies. Concentration of linker molecules was also independently verified by absorption spectroscopy using the highly collimated micro-beam light of Diamond B23 beamline. The detection was achieved through the binding reaction between the antibody and varying concentrations of Clusterin antigen from 1 to 100 pg/mL, as well as specificity tests using human chorionic gonadotropin (hCG), a glycoprotein risk biomarker of certain cancers. The GFETs were characterized using direct current (DC) 4-probe electrical resistance (4-PER) measurements, which demonstrated a limit of detection of the biosensors to be ∼ 300 fg/mL (4 fM). Comparison with back-gated Dirac voltage shifts with varying concentration of Clusterin show 4-PER measurements to be more accurate, at present, and point to a requirement for further optimisation of the fabrication processes for our next generation of GFET sensors. Thus, we have successfully fabricated a promising set of GFET biosensors for the detection of Clusterin protein biomarker. The developed GFET biosensors are entirely generic and also have the potential to be applied to a variety of other disease detection applications such as Parkinson's, cancer, and cardiovascular.

4.
Front Public Health ; 8: 574111, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33324597

RESUMO

Non-communicable diseases (NCDs) are of increasing concern for society and national governments, as well as globally due to their high mortality rate. The main risk factors of NCDs can be classified into the categories of self-management, genetic factors, environmental factors, factors of medical conditions, and socio-demographic factors. The main focus is on the elements of self-management and to reach a consensus about the influence of food on risk management and actions toward the prevention of NCDs at all stages of life. Nutrition interventions are essential in managing the risk of NCDs. As they are of the utmost importance, this review highlights NCDs and their risk factors and outlines several common prevention strategies. We foresee that the best prevention management strategy will include individual (lifestyle management), societal (awareness management), national (health policy decisions), and global (health strategy) elements, with target actions, such as multi-sectoral partnership, knowledge and information management, and innovations. The most effective preventative strategy is the one that leads to changes in lifestyle with respect to diet, physical activities, cessation of smoking, and the control of metabolic disorders.


Assuntos
Doenças não Transmissíveis , Dieta , Política de Saúde , Humanos , Estilo de Vida , Doenças não Transmissíveis/epidemiologia , Fatores de Risco
5.
Sci Rep ; 10(1): 19517, 2020 11 11.
Artigo em Inglês | MEDLINE | ID: mdl-33177577

RESUMO

In this study, synthetic polymeric particles were effectively fabricated by combining modern technologies of artificial intelligence (AI) and microfluidics. Because size uniformity is a key factor that significantly influences the stability of polymeric particles, therefore, this work aimed to establish a new AI application using machine learning technology for prediction of the size of poly(D,L-lactide-co-glycolide) (PLGA) microparticles produced by diverse microfluidic systems either in the form of single or multiple particles. Experimentally, the most effective factors for tuning droplet/particle sizes are PLGA concentrations and the flow rates of dispersed and aqueous phases in microfluidics. These factors were utilized to develop five different and simple in structure artificial neural network (ANN) models that are capable of predicting PLGA particle sizes produced by different microfluidic systems either individually or jointly merged. The systematic development of ANN models allowed ultimate construction of a single in silico model which consists of data for three different microfluidic systems. This ANN model eventually allowed rapid prediction of particle sizes produced using various microfluidic systems. This AI application offers a new platform for further rapid and economical exploration of polymer particles production in defined sizes for various applications including biomimetic studies, biomedicine, and pharmaceutics.

6.
Sensors (Basel) ; 20(6)2020 Mar 19.
Artigo em Inglês | MEDLINE | ID: mdl-32204503

RESUMO

Designing and development of electrochemical biosensors enable molecule sensing and quantification of biochemical compositions with multitudinous benefits such as monitoring, detection, and feedback for medical and biotechnological applications. Integrating bioinspired materials and electrochemical techniques promote specific, rapid, sensitive, and inexpensive biosensing platforms for (e.g., point-of-care testing). The selection of biomaterials to decorate a biosensor surface is a critical issue as it strongly affects selectivity and sensitivity. In this context, smart biomaterials with the intrinsic self-assemble capability like bacterial surface (S-) layer proteins are of paramount importance. Indeed, by forming a crystalline two-dimensional protein lattice on many sensors surfaces and interfaces, the S-layer lattice constitutes an immobilization matrix for small biomolecules and lipid membranes and a patterning structure with unsurpassed spatial distribution for sensing elements and bioreceptors. This review aims to highlight on exploiting S-layer proteins in biosensor technology for various applications ranging from detection of metal ions over small organic compounds to cells. Furthermore, enzymes immobilized on the S-layer proteins allow specific detection of several vital biomolecules. The special features of the S-layer protein lattice as part of the sensor architecture enhances surface functionalization and thus may feature an innovative class of electrochemical biosensors.


Assuntos
Técnicas Biossensoriais , Técnicas Eletroquímicas , Glicoproteínas de Membrana/isolamento & purificação , Humanos , Íons/química , Íons/isolamento & purificação , Glicoproteínas de Membrana/química , Metais/química , Metais/isolamento & purificação , Testes Imediatos
7.
Biomolecules ; 9(12)2019 12 17.
Artigo em Inglês | MEDLINE | ID: mdl-31861140

RESUMO

Recently more consideration has been given to the use of renewable materials and agricultural residues. Wheat production is increasing yearly and correspondingly, the volume of by-products from the wheat process is increasing, as well. It is important to find the use of the residuals for higher value-added products, and not just for the food industry or animal feed purposes as it is happening now. Agricultural residue of the roller milled wheat grain is a wheat bran description. The low-cost of wheat bran and its composition assortment provides a good source of substrate for various enzymes and organic acids production and other biotechnological applications. The main purpose of this review article is to look into recent trends, developments, and applications of wheat bran.


Assuntos
Fibras na Dieta , Animais , Biotecnologia , Fibras na Dieta/economia , Fibras na Dieta/metabolismo , Fibras na Dieta/uso terapêutico , Humanos
8.
Biomed Microdevices ; 21(3): 62, 2019 07 04.
Artigo em Inglês | MEDLINE | ID: mdl-31273471

RESUMO

Cells are the basic units of life, and can be mimicked to create artificial analogs enabling the investigation of cellular mechanisms under controlled conditions. Building biomimetic systems ranging from proto-cells to cell-like objects such as compartment membranes can be achieved by collecting biobricks that self-assemble to build simplified models performing specific functions. Hence, scientists can develop and optimize new synthetic cells with biological functions by taking inspiration from nature and exploiting the advantages of synthetic biology. However, the bottom-down approach is not restricted to the basic principles of biological cells, and new mimicry systems can be designed starting with a combination of living and non-living simple molecules to focus on a cellular machinery function. In recent years, microfluidic devices have been well established to engineer bioarchitecture models resembling cell-like structures involving vesicles, compartmentalization, synthetic membranes, and the chip itself as a synthetic cell. This review aims to highlight the role of biological cells and their impact on inspiring the development of biomimetic models. The combination of the principles of synthetic biology with microfluidic technology represents the newly-introduced field of synthetic cells and synthetic membranes that can be further exploited in diagnostic and therapeutic applications.


Assuntos
Biomimética/instrumentação , Dispositivos Lab-On-A-Chip , Desenho de Equipamento , Bicamadas Lipídicas/metabolismo
9.
Biophys Chem ; 251: 106178, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31102749

RESUMO

Development of synthetic bioarchitectures to improve our understanding of biological systems and produce biosynthetic models with new functions has attracted substantial interest. Synthetic HDL-like phospholipid nanodiscs are a relatively new model of nanoparticles that present a promising carrier for drug delivery and membrane protein investigations. Nanodiscs are soluble nanoscale phospholipid bilayers that are produced based on the self-assembly of phospholipids, membrane scaffold proteins (MSP) and an embedded peptide/protein of interest. To determine the effect of conjugating a protein with a probe, the model protein bovine serum albumin (BSA) with or without FITC conjugation was attached onto 100% 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-choline (POPC) nanodiscs. The generated discs were analyzed by Fast Protein Liquid Chromatography (FPLC), dynamic light scattering (DLS), and UV-VIS spectroscopy. Empty, BSA- and FITC-BSA-Nanodiscs exhibited different size, charge and elution characteristics as well as different release profiles. Thus, conjugation of proteins to be adsorbed onto nanodiscs surfaces with fluorophores can affect the physical and release properties of nanodiscs, thereby potentially impacting their biophysical, delivery and imaging applications.


Assuntos
Albuminas/química , Sistemas de Liberação de Medicamentos , Bicamadas Lipídicas/química , Nanoestruturas/química , Adsorção , Modelos Moleculares , Tamanho da Partícula , Propriedades de Superfície
10.
J Med Biochem ; 38(2): 145-152, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30867642

RESUMO

Background: Although vitamin D in not a traditional marker for cardiovascular and renal diseases, several studies have proposed a correlation between vitamin D deficiency and these diseases due to the effect of vitamin D on endothelial function. Asymmetric and symmetric dimethyl arginine (ADMA and SDMA, respectively) are endogenous markers of endothelial dysfunction, and are considered as future markers for the assessment of cardiovascular and renal diseases. The present study investigated the association of kidney function tests (urea and creatinine) and dimethylarginine toxins (ADMA and SDMA) in women with vitamin D insufficiency or deficiency. Indeed, sex hormones (estrogen and testosterone) were analyzed in the participants. Methods: Women were divided into two groups: premenopausal women (younger than 50 years) and postmenopausal women (older than 50 years). Urea, creatinine, estrogen, testosterone, ADMA, and SDMA levels were analyzed when vitamin D level was deficient or insufficient in the participants. Results: The premenopausal women group showed no significant correlations between dimethylarginine toxins and renal failure tests or sex hormones. In the elderly (postmenstrual) women group, only SDMA was significantly correlated with urea and creatinine, while both ADMA and SDMA were not correlated with sex hormones. Conclusions: Although ADMA and SDMA are promising candidates of endothelial dysfunction and are increased in menopause and aging, no direct link between ADMA and further progression of renal failure was observed in women with low vitamin D levels. In contrast, a possible direct correlation between SDMA and renal dysfunction was noticed, but only in an age-dependent manner.

11.
J Med Biochem ; 37(4): 441-447, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30584403

RESUMO

Background: Vitamin D deficiency has been reported to be associated with pregnancy loss. Asymmetric dimethyl-L-arginine (ADMA) and symmetric dimethyl-L-arginine (SDMA) are arginine analogues that have direct and indirect effects on nitric oxide (NO) synthesis and endothelial dysfunction. This study aimed to evaluate ADMA and SDMA levels among women with history of pregnancy loss compared to women without history of pregnancy loss and all participants were suffering from vitamin D deficiency. Methods: To investigate the relationship between vitamin D deficiency and ADMA and SDMA, both groups of women were experiencing vitamin D deficiency. All women enrolled in this study had a vitamin D level below 75 nmol/L and were not pregnant. ADMA and SDMA levels were investigated in 28 women without a history of pregnancy loss and 19 women with a history of pregnancy loss. Results: No statistically significant differences were found in ADMA and SDMA levels among the two groups. The correlation analysis showed that vitamin D deficiency was not significantly inversely correlated with ADMA and SDMA in women without a history of pregnancy loss, but was significantly correlated with SDMA in women with a history of pregnancy loss. Conclusions: Vitamin D deficiency, in women with or without a history of failed clinical pregnancies, has no effect on the circulating levels of ADMA and SDMA. Further studies are needed to investigate any possible link between these parameters.

12.
Genes (Basel) ; 9(6)2018 Jun 06.
Artigo em Inglês | MEDLINE | ID: mdl-29882823

RESUMO

Understanding the mechanisms that govern nervous tissues function remains a challenge. In vitro two-dimensional (2D) cell culture systems provide a simplistic platform to evaluate systematic investigations but often result in unreliable responses that cannot be translated to pathophysiological settings. Recently, microplatforms have emerged to provide a better approximation of the in vivo scenario with better control over the microenvironment, stimuli and structure. Advances in biomaterials enable the construction of three-dimensional (3D) scaffolds, which combined with microfabrication, allow enhanced biomimicry through precise control of the architecture, cell positioning, fluid flows and electrochemical stimuli. This manuscript reviews, compares and contrasts advances in nervous tissues-on-a-chip models and their applications in neural physiology and disease. Microplatforms used for neuro-glia interactions, neuromuscular junctions (NMJs), blood-brain barrier (BBB) and studies on brain cancer, metastasis and neurodegenerative diseases are addressed. Finally, we highlight challenges that can be addressed with interdisciplinary efforts to achieve a higher degree of biomimicry. Nervous tissue microplatforms provide a powerful tool that is destined to provide a better understanding of neural health and disease.

13.
Genes (Basel) ; 9(3)2018 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-29509709

RESUMO

Synthetic biology is a rapidly growing multidisciplinary branch of science which aims to mimic complex biological systems by creating similar forms. Constructing an artificial system requires optimization at the gene and protein levels to allow the formation of entire biological pathways. Advances in cell-free synthetic biology have helped in discovering new genes, proteins, and pathways bypassing the complexity of the complex pathway interactions in living cells. Furthermore, this method is cost- and time-effective with access to the cellular protein factory without the membrane boundaries. The freedom of design, full automation, and mimicking of in vivo systems reveal advantages of synthetic biology that can improve the molecular understanding of processes, relevant for life science applications. In parallel, in vitro approaches have enhanced our understanding of the living system. This review highlights the recent evolution of cell-free gene design, proteins, and cells integrated with microfluidic platforms as a promising technology, which has allowed for the transformation of the concept of bioprocesses. Although several challenges remain, the manipulation of biological synthetic machinery in microfluidic devices as suitable 'homes' for in vitro protein synthesis has been proposed as a pioneering approach for the development of new platforms, relevant in biomedical and diagnostic contexts towards even the sensing and monitoring of environmental issues.

14.
Genes (Basel) ; 9(2)2018 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-29443890

RESUMO

Hepatic oval cells (HOCs) are considered the progeny of the intrahepatic stem cells that are found in a small population in the liver after hepatocyte proliferation is inhibited. Due to their small number, isolation and capture of these cells constitute a challenging task for immunosensor technology. This work describes the development of a 3D-printed continuous flow system and exploits disposable screen-printed electrodes for the rapid detection of HOCs that over-express the OV6 marker on their membrane. Multiwall carbon nanotube (MWCNT) electrodes have a chitosan film that serves as a scaffold for the immobilization of oval cell marker antibodies (anti-OV6-Ab), which enhance the sensitivity of the biomarker and makes the designed sensor specific for oval cells. The developed sensor can be easily embedded into the 3D-printed flow cell to allow cells to be exposed continuously to the functionalized surface. The continuous flow is intended to increase capture of most of the target cells in the specimen. Contact angle measurements were performed to characterize the nature and quality of the modified sensor surface, and electrochemical measurements (cyclic voltammetry (CV) and square wave voltammetry (SWV)) were performed to confirm the efficiency and selectivity of the fabricated sensor to detect HOCs. The proposed method is valuable for capturing rare cells and could provide an effective tool for cancer diagnosis and detection.

15.
Genes (Basel) ; 9(2)2018 Feb 16.
Artigo em Inglês | MEDLINE | ID: mdl-29462948

RESUMO

Microfluidic devices present unique advantages for the development of efficient drug carrier particles, cell-free protein synthesis systems, and rapid techniques for direct drug screening. Compared to bulk methods, by efficiently controlling the geometries of the fabricated chip and the flow rates of multiphase fluids, microfluidic technology enables the generation of highly stable, uniform, monodispersed particles with higher encapsulation efficiency. Since the existing preclinical models are inefficient drug screens for predicting clinical outcomes, microfluidic platforms might offer a more rapid and cost-effective alternative. Compared to 2D cell culture systems and in vivo animal models, microfluidic 3D platforms mimic the in vivo cell systems in a simple, inexpensive manner, which allows high throughput and multiplexed drug screening at the cell, organ, and whole-body levels. In this review, the generation of appropriate drug or gene carriers including different particle types using different configurations of microfluidic devices is highlighted. Additionally, this paper discusses the emergence of fabricated microfluidic cell-free protein synthesis systems for potential use at point of care as well as cell-, organ-, and human-on-a-chip models as smart, sensitive, and reproducible platforms, allowing the investigation of the effects of drugs under conditions imitating the biological system.

16.
Diagnostics (Basel) ; 8(1)2018 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-29316718

RESUMO

We report on the development of label-free chemical vapour deposition (CVD) graphene field effect transistor (GFET) immunosensors for the sensitive detection of Human Chorionic Gonadotropin (hCG), a glycoprotein risk biomarker of certain cancers. The GFET sensors were fabricated on Si/SiO2 substrate using photolithography with evaporated chromium and sputtered gold contacts. GFET channels were functionalised with a linker molecule to an immobile anti-hCG antibody on the surface of graphene. The binding reaction of the antibody with varying concentration levels of hCG antigen demonstrated the limit of detection of the GFET sensors to be below 1 pg/mL using four-probe electrical measurements. We also show that annealing can significantly improve the carrier transport properties of GFETs and shift the Dirac point (Fermi level) with reduced p-doping in back-gated measurements. The developed GFET biosensors are generic and could find applications in a broad range of medical diagnostics in addition to cancer, such as neurodegenerative (Alzheimer's and Parkinson's ) and cardiovascular disorders.

17.
Sensors (Basel) ; 18(1)2018 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-29361679

RESUMO

Clusterin (CLU) has been associated with the clinical progression of Alzheimer's disease (AD) and described as a potential AD biomarker in blood plasma. Due to the enormous attention given to cerebrospinal fluid (CSF) biomarkers for the past couple of decades, recently found blood-based AD biomarkers like CLU have not yet been reported for biosensors. Herein, we report the electrochemical detection of CLU for the first time using a screen-printed carbon electrode (SPCE) modified with 1-pyrenebutyric acid N-hydroxysuccinimide ester (Pyr-NHS) and decorated with specific anti-CLU antibody fragments. This bifunctional linker molecule contains succinylimide ester to bind protein at one end while its pyrene moiety attaches to the carbon surface by means of π-π stacking. Cyclic voltammetric and square wave voltammetric studies showed the limit of detection down to 1 pg/mL and a linear concentration range of 1-100 pg/mL with good sensitivity. Detection of CLU in spiked human plasma was demonstrated with satisfactory recovery percentages to that of the calibration data. The proposed method facilitates the cost-effective and viable production of label-free point-of-care devices for the clinical diagnosis of AD.


Assuntos
Clusterina/análise , Doença de Alzheimer , Biomarcadores , Técnicas Biossensoriais , Eletrodos , Humanos , Limite de Detecção , Pirenos
18.
Genes (Basel) ; 8(10)2017 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-28953246

RESUMO

Air quality depends on the various gases and particles present in it. Both natural phenomena and human activities affect the cleanliness of air. In the last decade, many countries experienced an unprecedented industrial growth, resulting in changing air quality values, and correspondingly, affecting our life quality. Air quality can be accessed by employing microchips that qualitatively and quantitatively determine the present gases and dust particles. The so-called particular matter 2.5 (PM2.5) values are of high importance, as such small particles can penetrate the human lung barrier and enter the blood system. There are cancer cases related to many air pollutants, and especially to PM2.5, contributing to exploding costs within the healthcare system. We focus on various current and potential future air pollutants, and propose solutions on how to protect our health against such dangerous substances. Recent developments in the Organ-on-Chip (OoC) technology can be used to study air pollution as well. OoC allows determination of pollutant toxicity and speeds up the development of novel pharmaceutical drugs.

19.
Biosens Bioelectron ; 94: 500-506, 2017 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-28343102

RESUMO

This study presents an efficient acoustic and hybrid three-dimensional (3D)-printed electrochemical biosensors for the detection of liver cancer cells. The biosensors function by recognizing the highly expressed tumor marker CD133, which is located on the surface of liver cancer cells. Detection was achieved by recrystallizing a recombinant S-layer fusion protein (rSbpA/ZZ) on the surface of the sensors. The fused ZZ-domain enables immobilization of the anti-CD133 antibody in a defined manner. These highly accessible anti-CD133 antibodies were employed as a sensing layer, thereby enabling the efficient detection of liver cancer cells (HepG2). The recognition of HepG2 cells was investigated in situ using a quartz crystal microbalance with dissipation monitoring (QCM-D), which enabled the label-free, real-time detection of living cells on the modified sensor surface under controlled conditions. Furthermore, the hybrid 3D additive printing strategy for biosensors facilitates both rapid development and small-scale manufacturing. The hybrid strategy of combining 3D-printed parts and more traditionally fabricated parts enables the use of optimal materials: a ceramic substrate with noble metals for the sensing element and 3D-printed capillary channels to guide and constrain the clinical sample. Cyclic voltammetry (CV) measurements confirmed the efficiency of the fabricated sensors. Most importantly, these sensors offer low-cost and disposable detection platforms for real-world applications. Thus, as demonstrated in this study, both fabricated acoustic and electrochemical sensing platforms can detect cancer cells and therefore may have further potential in other clinical applications and drug-screening studies.


Assuntos
Antígeno AC133/isolamento & purificação , Técnicas Biossensoriais , Neoplasias Hepáticas/diagnóstico , Antígeno AC133/química , Acústica , Técnicas Eletroquímicas , Células Hep G2 , Humanos , Neoplasias Hepáticas/genética , Impressão Tridimensional , Técnicas de Microbalança de Cristal de Quartzo
20.
Biomater Sci ; 3(10): 1406-13, 2015 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-26236783

RESUMO

We present an elegant synthesis and reconstitution approach for functional studies of voltage responsive membrane proteins. For such studies, we propose a planar architecture of an S-layer-supported lipid membrane as a suitable matrix for presenting unmodified membrane protein species, and here we focus on the voltage-dependent anion channel (VDAC) from human mitochondria. The presented cell-free strategy, in which VDAC proteins are synthesized in bacterial cell lysate, into a membrane structure, offers a great advantage in the study of such subtle membrane proteins over the conventional, cell-based synthesis approach in terms of reproducibility. The material-assay combination is superior over cell-culture related synthesis and purification approaches as here we bypass by a one-step synthesis procedure the complex cell culture, and expression and purification endeavours, and, moreover, the protein of interest never has to be detergent solubilized and had been synthesized de novo. We provide here a detailed description from the all over procedure and our first results, describing in detail the cell-free synthesis and robustness of such a material-assay combination: functional VDAC protein species embedded in a planar membrane architecture and ready for electrochemical characterization.


Assuntos
Bicamadas Lipídicas/química , Lipídeos/química , Proteínas de Membrana/química , Mitocôndrias/química , Ribossomos/química , Canais de Ânion Dependentes de Voltagem/síntese química , Técnicas Eletroquímicas , Humanos , Imidas/química , Bicamadas Lipídicas/metabolismo , Proteínas de Membrana/metabolismo , Mitocôndrias/metabolismo , Fosfatidiletanolaminas/química , Propilaminas/química , Canais de Ânion Dependentes de Voltagem/química , Canais de Ânion Dependentes de Voltagem/metabolismo
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