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Towards Nanoscale Biomedical Devices in Medicine: Biofunctional and Spectroscopic Characterization of Superparamagnetic Nanoparticles

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Abstract

Medical interest in nanotechnology originates from a belief that nanoscale therapeutic devices can be constructed and directed towards its target inside the human body. Such nanodevices can be engineered by coupling superparamagnetic nanoparticle to biomedically active proteins. We hereby report the immobilization of a PhEst, a S-formylglutathione hydrolase from the psychrophilic P. haloplanktis TAC125 onto the gold coated surface of modified superparamagnetic core-shell nanoparticles (Fe3O4@Au). The synthesis of the nanoparticles is also reported. S-formylglutathione hydrolases constitute a family of ubiquitous enzymes which play a key role in formaldehyde detoxification both in prokaryotes and eukaryotes. PhEst was originally annotated as a putative feruloyl esterase, an enzyme that releases ferulic acid (an antioxidant reactive towards free radicals such as reactive oxygen species) from polysaccharides esters. Dynamic light scattering, scanning electron microscopy with energy dispersive X-ray spectroscopy, UV–visible absorption spectroscopy, fluorescence spectroscopy, magnetic separation technique and enzyme catalytic assay confirmed the chemical composition of the gold covered superparamagnetic nanoparticles, the binding and activity of the enzyme onto the nanoparticles. Activity data in U/ml confirmed that the immobilized enzyme is approximately 2 times more active than the free enzyme in solution. Such particles can be directed with external magnetic fields for bio-separation and focused towards a medical target for therapeutical as well as bio-sensor applications.

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Abbreviations

BSA:

Bovine serum albumin

DLS:

Dynamic light scattering

DPPH:

1,1-diphenyl-2picrylhydrazyl

EDS:

Energy dispersive X-ray spectroscopy

Fe3O4@Au:

Superparamagnetic core(magnetite)-shell(gold) nanoparticles

MRI:

Magnetic resonance imaging

PhEst:

Putative feruloyl esterase

pNPAc:

p-nitro phenyl acetate

ROS:

Reactive oxygen species

SEM:

Scanning electron microscopy

U/ml:

Unit/milliliter (1 unit U is the amount of enzyme that catalyses the reaction of 1 μmol of substrate per minute)

UV–visible:

Ultraviolet–visible

FWHM:

Full width at half maximum

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Acknowledgments

The authors are indebted to Dr. Sabato D’Auria and Vincenzo Aurialia from Institute of Protein Biochemistry, CNR, Via Pietro Castellino, 111, 80131 Napoli, Italy for providing the enzyme.

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Authors and Affiliations

  1. NanoBiotechnology Group, Department of Physics and Nanotechnology, Aalborg University, Skjernvej 4A, Aalborg, Denmark

    Antonietta Parracino, Gnana Prakash Gajula, Ane Kold di Gennaro, Maria Teresa Neves-Petersen & Steffen B. Petersen

  2. Department of Physics and Nanotechnology, Aalborg University, Skjernvej 4A, Aalborg, Denmark

    Jens Rafaelsen

  3. The Institute for Lasers, Photonics and Biophotonics, University at Buffalo, The State University of New York Buffalo, Buffalo, NY, 14260-3000, USA

    Steffen B. Petersen

Authors
  1. Antonietta Parracino
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  2. Gnana Prakash Gajula
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  3. Ane Kold di Gennaro
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  4. Maria Teresa Neves-Petersen
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  5. Jens Rafaelsen
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  6. Steffen B. Petersen
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Corresponding author

Correspondence to Steffen B. Petersen.

Additional information

Antonietta Parracino and Gnana Prakash Gajula contributed equally to the paper.

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Parracino, A., Gajula, G.P., di Gennaro, A.K. et al. Towards Nanoscale Biomedical Devices in Medicine: Biofunctional and Spectroscopic Characterization of Superparamagnetic Nanoparticles. J Fluoresc 21, 663–672 (2011). https://doi.org/10.1007/s10895-010-0754-6

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  • DOI: https://doi.org/10.1007/s10895-010-0754-6

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