Calorimetric Sensor for Ethanol Using Ni2+-nitrilotriacetic Acid (NTA) Resin Immobilized Alcohol Dehydrogenase (ADH)

Background: A simple, fast and economic analytical method for the determination of ethanol is important for clinical, biological, forensic and physico-legal purposes. Methods: Ni2+-NTA resin was used as an immobilization matrix for the simple one-step purification/ immobilization of his6-tagged ADH. Different alcohols with a concentration range of 0.5-50% V/V, namely methanol, ethanol and propanol were measured using prepared ADH enzyme thermistor. The ethanol content of Tsingtao beer was tested as a real sample containing alcohol. Reproducibility and stability of prepared ADH enzyme thermistor were also investigated by repeated measurements. Results: In comparison to the controlled pore glass (a common used support for the immobilization of enzyme) used in thermal biosensor, the use of Ni2+-NTA resin not only led to simple one-step purification/ immobilization by his6-tagged ADH binding to Ni2+-NTA resin, but also made the immobilizing supports reusable. The prepared biosensor can be used to determine ethanol and methanol by the calorimetric measurement. A linear range of 1 -32% (V/V) and 2-20% (V/V) was observed for ethanol and methanol, respectively. The detection limits were 0.3% (V/V) and 1% (V/V) for ethanol and methanol, respectively. The tested ethanol concentration of Tsingtao beer was 4.5% V/V, which is comparable with the labeled alcohol by volume (ABV) 4.80%. Conclusion: Ni2+-NTA resin, as an immobilization matrix in ET sensor, provides a simple one-step purification/immobilization for His6-tagged recombinase and a reusable immobilization matrix. The prepared biosensor exhibits good repeatability and stability. Such a new biosensor shows great promise for rapid, simple, and cost-effective analysis of ethanol and methanol, both in qualitative and in quantitative tests.

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Parameters Controlling the One-Step Derivatization of Controlled Pore Glass with a Diol for Solid-Phase Synthesis of 3‘-Modified Oligonucleotides

Organic Process Research & Development ◽

10.1021/op050221v ◽

2006 ◽

Vol 10 (3) ◽

pp. 403-408 ◽

Cited By ~ 2

Author(s):

Alain Laurent ◽

Carole Chaix

Keyword(s):

Solid Phase Synthesis ◽

Solid Phase ◽

Modified Oligonucleotides ◽

Phase Synthesis ◽

Controlled Pore Glass ◽

Pore Glass ◽

One Step ◽

The One

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A one step derivatization of controlled pore glass for oligonucleotide solid-phase synthesis

Tetrahedron Letters ◽

10.1016/j.tetlet.2004.09.167 ◽

2004 ◽

Vol 45 (48) ◽

pp. 8883-8887 ◽

Cited By ~ 4

Author(s):

Alain Laurent ◽

Bertrand de Lambert ◽

Marie-Thérèse Charreyre ◽

Bernard Mandrand ◽

Carole Chaix

Keyword(s):

Solid Phase Synthesis ◽

Solid Phase ◽

Phase Synthesis ◽

Controlled Pore Glass ◽

Pore Glass ◽

One Step

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Nitrilotriacetic Acid (NTA) Resin as a Reversible Immobilization Matrix for Polyhistidine Tagged Enzyme in Enzyme Thermistor

Sensor Letters ◽

10.1166/sl.2009.1236 ◽

2009 ◽

Vol 7 (6) ◽

pp. 1072-1076 ◽

Cited By ~ 2

Author(s):

YongJin Li ◽

HaoYong Wang ◽

WeiHong Xie

Keyword(s):

Nitrilotriacetic Acid ◽

Reversible Immobilization ◽

Immobilization Matrix ◽

Enzyme Thermistor

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Antithrombotic Therapy for DVT/PE

Hämostaseologie ◽

10.1055/s-0038-1659983 ◽

1997 ◽

Vol 17 (03) ◽

pp. 161-162

Author(s):

Thomas Hyers

Keyword(s):

Molecular Weight ◽

Low Molecular Weight Heparin ◽

Oral Anticoagulants ◽

Cost Effective ◽

Therapeutic Agents ◽

Great Promise ◽

Term Therapy ◽

Low Molecular Weight ◽

Long Term Therapy

SummaryProblems with unfractionated heparin as an antithrombotic have led to the development of new therapeutic agents. Of these, low molecular weight heparin shows great promise and has led to out-patient therapy of DVT/PE in selected patients. Oral anticoagulants remain the choice for long-term therapy. More cost-effective ways to give oral anticoagulants are needed.

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Improved Conditions for Solid Phase Synthesis of Oligonucleotides on PS-PEG Copolymers

Zeitschrift für Naturforschung B ◽

10.1515/znb-1995-0720 ◽

1995 ◽

Vol 50 (7) ◽

pp. 1096-1100 ◽

Cited By ~ 14

Author(s):

Ernst Bayer ◽

Konrad Bleicher ◽

Martin Maier

Keyword(s):

Electrospray Mass Spectrometry ◽

Solid Phase Synthesis ◽

Solid Phase ◽

Chemical Properties ◽

Oligonucleotide Synthesis ◽

Phase Synthesis ◽

Controlled Pore Glass ◽

Product Identification ◽

Pore Glass ◽

And Chemical Properties

Polystyrene-polyethylene glycol (PS-PEG) tentacle polymers with loadings of up to 60/<μmol/g were used for standard oligonucleotide synthesis. As these resins are easy to handle and stable under reaction and cleavage conditions they may be used alternatively to controlled pore glass (CPG) as the most commonly used solid support for oligonucleotide synthesis. However, structural and chemical properties of the PS-PEG resins require modified conditions to guarantee syntheses with high coupling efficiencies. Oligonucleotides (ODN ) of various sequences and lengths have successfully been synthesized using HPLC and capillary electrophoresis (CE) for purity control. Additionally, electrospray mass spectrometry (ES-MS) was used for product identification.

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Novel Regeneration Approach for Creating Reusable FO-SPR Probes with NTA Surface Chemistry

Nanomaterials ◽

10.3390/nano11010186 ◽

2021 ◽

Vol 11 (1) ◽

pp. 186

Author(s):

Jia-Huan Qu ◽

Karen Leirs ◽

Remei Escudero ◽

Žiga Strmšek ◽

Roman Jerala ◽

...

Keyword(s):

Surface Chemistry ◽

Fluorescent Protein ◽

Cost Effective ◽

Nitrilotriacetic Acid ◽

Antibody Fragments ◽

Rapid Screening ◽

Red Fluorescent Protein ◽

Effective Manner ◽

Surface Regeneration ◽

Sensitivity Specificity

To date, surface plasmon resonance (SPR) biosensors have been exploited in numerous different contexts while continuously pushing boundaries in terms of improved sensitivity, specificity, portability and reusability. The latter has attracted attention as a viable alternative to disposable biosensors, also offering prospects for rapid screening of biomolecules or biomolecular interactions. In this context here, we developed an approach to successfully regenerate a fiber-optic (FO)-SPR surface when utilizing cobalt (II)-nitrilotriacetic acid (NTA) surface chemistry. To achieve this, we tested multiple regeneration conditions that can disrupt the NTA chelate on a surface fully saturated with His6-tagged antibody fragments (scFv-33H1F7) over ten regeneration cycles. The best surface regeneration was obtained when combining 100 mM EDTA, 500 mM imidazole and 0.5% SDS at pH 8.0 for 1 min with shaking at 150 rpm followed by washing with 0.5 M NaOH for 3 min. The true versatility of the established approach was proven by regenerating the NTA surface for ten cycles with three other model system bioreceptors, different in their size and structure: His6-tagged SARS-CoV-2 spike fragment (receptor binding domain, RBD), a red fluorescent protein (RFP) and protein origami carrying 4 RFPs (Tet12SN-RRRR). Enabling the removal of His6-tagged bioreceptors from NTA surfaces in a fast and cost-effective manner can have broad applications, spanning from the development of biosensors and various biopharmaceutical analyses to the synthesis of novel biomaterials.

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Electrochemical Response of Glucose Oxidase Adsorbed on Laser-Induced Graphene

Nanomaterials ◽

10.3390/nano11081893 ◽

2021 ◽

Vol 11 (8) ◽

pp. 1893

Author(s):

Sónia O. Pereira ◽

Nuno F. Santos ◽

Alexandre F. Carvalho ◽

António J. S. Fernandes ◽

Florinda M. Costa

Keyword(s):

Electron Transfer ◽

Glucose Oxidase ◽

Ph Dependence ◽

High Surface ◽

High Surface Area ◽

Cost Effective ◽

Great Promise ◽

Glucose Detection ◽

Half Wave ◽

Electrochemical Transducers

Carbon-based electrodes have demonstrated great promise as electrochemical transducers in the development of biosensors. More recently, laser-induced graphene (LIG), a graphene derivative, appears as a great candidate due to its superior electron transfer characteristics, high surface area and simplicity in its synthesis. The continuous interest in the development of cost-effective, more stable and reliable biosensors for glucose detection make them the most studied and explored within the academic and industry community. In this work, the electrochemistry of glucose oxidase (GOx) adsorbed on LIG electrodes is studied in detail. In addition to the well-known electroactivity of free flavin adenine dinucleotide (FAD), the cofactor of GOx, at the expected half-wave potential of −0.490 V vs. Ag/AgCl (1 M KCl), a new well-defined redox pair at 0.155 V is observed and shown to be related to LIG/GOx interaction. A systematic study was undertaken in order to understand the origin of this activity, including scan rate and pH dependence, along with glucose detection tests. Two protons and two electrons are involved in this reaction, which is shown to be sensitive to the concentration of glucose, restraining its origin to the electron transfer from FAD in the active site of GOx to the electrode via direct or mediated by quinone derivatives acting as mediators.

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3D Printed Microfluidic Devices for Drug Release Assays

Pharmaceutics ◽

10.3390/pharmaceutics13010013 ◽

2020 ◽

Vol 13 (1) ◽

pp. 13

Author(s):

Benzion Amoyav ◽

Yoel Goldstein ◽

Eliana Steinberg ◽

Ofra Benny

Keyword(s):

3D Printing ◽

Drug Release ◽

Particle Production ◽

Cost Effective ◽

Drug Dissolution ◽

Cytotoxicity Test ◽

High Durability ◽

One Step ◽

Cost Effective Alternative ◽

Multiple Recycling

Microfluidics research for various applications, including drug delivery, cell-based assays and biomedical research has grown exponentially. Despite this technology’s enormous potential, drawbacks include the need for multistep fabrication, typically with lithography. We present a one-step fabrication process of a microfluidic chip for drug dissolution assays based on a 3D printing technology. Doxorubicin porous and non-porous microspheres, with a mean diameter of 250µm, were fabricated using a conventional “batch” or microfluidic method, based on an optimized solid-in-oil-in-water protocol. Microspheres fabricated with microfluidics system exhibited higher encapsulation efficiency and drug content as compared with batch formulations. We determined drug release profiles of microspheres in varying pH conditions using two distinct dissolution devices that differed in their mechanical barrier structures. The release profile of the “V” shape barrier was similar to that of the dialysis sac test and differed from the “basket” barrier design. Importantly, a cytotoxicity test confirmed biocompatibility of the printed resin. Finally, the chip exhibited high durability and stability, enabling multiple recycling sessions. We show how the combination of microfluidics and 3D printing can reduce costs and time, providing an efficient platform for particle production while offering a feasible cost-effective alternative to clean-room facility polydimethylsiloxane-based chip microfabrication.

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