Evaluation of Immobilized Hfmo3 on Magnetic Nanoparticles by Capillary Zone Electrophoresis

Ramana, Pranov; Herman, Ward; Hiroux, Charlotte; Adams, Erwin; Augustijns, Patrick; Schepdael, Ann Van

doi:10.4155/bio-2016-0151

Cited by 7 publications

(16 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Immobilization of hFMO3 on magnetic nanoparticles has been discussed in detail by Ramana et al . . hFMO3 was also immobilized on electrodes by Castrignanò et al .…”

Section: Resultsmentioning

confidence: 99%

“…In order to demonstrate the trapping efficiency and the ability to perform an in‐line reaction, hFMO3 was immobilized on amino functionalized MMs based on protocols previously described . For all washing steps performed off‐line, a MM‐separator® M12+12 (MagnaMedics Diagnostics B.V.) was used.…”

Section: Methodsmentioning

confidence: 99%

“…Off‐line reactions were performed as previously described in the literature . Free hFMO3 (0.075 mg/mL final concentration) or hFMO3 (0.075 mg/mL) immobilized on MMs was added to the incubation buffer to which 5 μL of 100 mM (5 mM final concentration) freshly dissolved MgCl 2 was added . NADPH was freshly prepared (2 mM final concentration) and added along with clozapine to the mixture that was incubated for 30 min at 37°C, 1500 RPM (total reaction volume = 100 μL).…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

An improved design to capture magnetic microparticles for capillary electrophoresis based immobilized microenzyme reactors

et al. 2018

Self Cite

View full text Add to dashboard Cite

In this paper, we demonstrate the effectiveness of a new 3D printed magnet holder that enables capture of magnetic microparticles in commercially available capillary electrophoresis equipment with a liquid or air based coolant system. The design as well as the method to capture magnetic microparticles inside the capillary are discussed. This setup was tested at temperature and pH values suitable for performing enzymatic reactions. To demonstrate its applicability in CE‐ immobilized microenzyme reactors (IMER) development, human flavin‐containing monooxygenase 3 and bovine serum albumin were immobilized on amino functionalized magnetic microparticles using glutaraldehyde. These microparticles were subsequently used to perform in‐line capillary electrophoresis with clozapine as a model substrate. This setup could be used further to establish CE‐IMERs of other drug metabolic enzymes in a commercially available liquid based capillary coolant system. The CE‐IMER setup was successful, although a subsequent decrease in enzyme activity was observed on repeated runs.

show abstract

“…Immobilization of hFMO3 on magnetic nanoparticles has been discussed in detail by Ramana et al . . hFMO3 was also immobilized on electrodes by Castrignanò et al .…”

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

An improved design to capture magnetic microparticles for capillary electrophoresis based immobilized microenzyme reactors

et al. 2018

Self Cite

View full text Add to dashboard Cite

show abstract

“…Similarly, intensive research on an hFMO3‐based IMER is being carried out by Ramana et al. in the group of Van Schepdael . Flavin‐containing monooxygenases (FMOs) are contributors to oxidative xenobiotic metabolism together with the well‐known CYP family.…”

Section: Applications Based On Pharmaceutical Relevancementioning

confidence: 99%

“…The particles with silica shell bore carboxyl moieties, thus the immobilization chemistry using carbodiimide as the activation agent was chosen. With this approach, we were able to reuse the constructed IMER for up to 20 runs, but with considerable loss of its [43,63]. Flavin-containing monooxygenases (FMOs) are contributors to oxidative xenobiotic metabolism together with the well-known CYP family.…”

Section: Drug-metabolizing Enzymesmentioning

confidence: 99%

Immobilized‐enzyme reactors integrated with capillary electrophoresis for pharmaceutical research

Schejbal

Glatz

2017

J of Separation Science

View full text Add to dashboard Cite

Enzymes play an essential role in many aspects of pharmaceutical research as drug targets, drug metabolizers, enzyme drugs and more. In this specific field, enzyme assays are required to meet a number of specific requirements, such as low cost, easy automation, and high reliability. The integration of an immobilized-enzyme reactor to capillary electrophoresis represents a unique approach to fulfilling these criteria by combining the benefits of enzyme immobilization, that is, increased stability and repeated use, as well as the minute sample consumption, short analysis time, and efficient analysis provided by capillary electrophoresis. In this review, we summarize, analyze, and discuss published works where pharmaceutically relevant enzymes were used to prepare capillary electrophoresis-integrated immobilized-enzyme reactors in an online manner. The presented assays are divided into three distinct groups based on the drug-enzyme relationship. The first, more extensively studied group employs enzymes that are considered to be therapeutic targets, the second group of assays present tools to assess drug metabolism and the third group assesses enzyme drugs. Furthermore, we examine various methods of enzyme immobilization and their implications for assay properties.

show abstract

Oxidoreductase enzymes: Characteristics, applications, and challenges as a biocatalyst

Cárdenas‐Moreno,

González‐Bacerio,

García Arellano

et al. 2023

Biotech and App Biochem

View full text Add to dashboard Cite

Oxidoreductases are enzymes with distinctive characteristics that favor their use in different areas, such as agriculture, environmental management, medicine, and analytical chemistry. Among these enzymes, oxidases, dehydrogenases, peroxidases, and oxygenases are very interesting. Because their substrate diversity, they can be used in different biocatalytic processes by homogeneous and heterogeneous catalysis. Immobilization of these enzymes has favored their use in the solution of different biotechnological problems, with a notable increase in the study and optimization of this technology in the last years. In this review, the main structural and catalytical features of oxidoreductases, their substrate specificity, immobilization, and usage in biocatalytic processes, such as bioconversion, bioremediation, and biosensors obtainment, are presented.

show abstract

Evaluation of Immobilized Hfmo3 on Magnetic Nanoparticles by Capillary Zone Electrophoresis

Abstract: A method to immobilize hFMO3 on magnetic nanoparticles has been described and evaluated in terms of enzyme activity, inhibition, pH stability and reusability.

Cited by 7 publications

References 20 publications

An improved design to capture magnetic microparticles for capillary electrophoresis based immobilized microenzyme reactors

An improved design to capture magnetic microparticles for capillary electrophoresis based immobilized microenzyme reactors

Immobilized‐enzyme reactors integrated with capillary electrophoresis for pharmaceutical research

Oxidoreductase enzymes: Characteristics, applications, and challenges as a biocatalyst

Contact Info

Product

Resources

About