About: Avidin Functionalized Maghernite Nanoparticles and Their Application for Recombinant Human Biotinyl-SERCA Purification     Goto   Sponge   Distinct   Permalink

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  • We report on the surface characterization, functionalization, and application of stable water suspensions of novel surface active maghemite narioparticles (SAMNs), characterized by a diameter of 11 +/- 2 nm and possessing peculiar colloidal properties and surface interactions. These features permitted the acquisition of titration curves and aqueous UV-vis spectra and suggested a role played by surface under-coordinated iron atoms. This new class of nanoparticles was obtained through an easy, inexpensive, one-step, green procedure and functionalized with ligands of high biotechnological interest, such as biotin and avidin, by simple incubation in aqueous solution. Bound avidin was determined by measuring the disappearance of free avidin absorbance at 280 nm, as a function of increasing nanoparticle concentration, showing the presence of 10 +/- 3 avidin molecules per nanoparticle. The biological activity of the SAMN@avidin complex was evaluated and the number of available biotin binding sites was determined, using biotinyl-fluorescein as a probe, showing that each bound avidin molecule is able to bind 2.8 +/- 0.8 biotin molecules, confirming the maintenance of biological activity and excellent binding capacity of the SAMN@avidin complex. Furthermore a Langmuir isotherm model was used to describe the biomolecule specific monolayer adsorption onto the particle surface, and in the case of avidin, the maximum adsorption capacity was 100 +/- 27 mu g avidin/mg SAMN, whereas the binding constant is 45.18 mu L mu g(-1). The SAMN@avidin complex was characterized by UV-vis spectroscopy, quartz crystal microbalance, FTIR spectroscopy, and transmission electron microscopy. Finally, SAMN@avidin was applied for the large scale purification of recombinant biotinylated human sarco/endoplasmic reticulum Ca2+-ATPase (hSERCA-2a), expressed by Saccharomyces cerevisiae.
  • We report on the surface characterization, functionalization, and application of stable water suspensions of novel surface active maghemite narioparticles (SAMNs), characterized by a diameter of 11 +/- 2 nm and possessing peculiar colloidal properties and surface interactions. These features permitted the acquisition of titration curves and aqueous UV-vis spectra and suggested a role played by surface under-coordinated iron atoms. This new class of nanoparticles was obtained through an easy, inexpensive, one-step, green procedure and functionalized with ligands of high biotechnological interest, such as biotin and avidin, by simple incubation in aqueous solution. Bound avidin was determined by measuring the disappearance of free avidin absorbance at 280 nm, as a function of increasing nanoparticle concentration, showing the presence of 10 +/- 3 avidin molecules per nanoparticle. The biological activity of the SAMN@avidin complex was evaluated and the number of available biotin binding sites was determined, using biotinyl-fluorescein as a probe, showing that each bound avidin molecule is able to bind 2.8 +/- 0.8 biotin molecules, confirming the maintenance of biological activity and excellent binding capacity of the SAMN@avidin complex. Furthermore a Langmuir isotherm model was used to describe the biomolecule specific monolayer adsorption onto the particle surface, and in the case of avidin, the maximum adsorption capacity was 100 +/- 27 mu g avidin/mg SAMN, whereas the binding constant is 45.18 mu L mu g(-1). The SAMN@avidin complex was characterized by UV-vis spectroscopy, quartz crystal microbalance, FTIR spectroscopy, and transmission electron microscopy. Finally, SAMN@avidin was applied for the large scale purification of recombinant biotinylated human sarco/endoplasmic reticulum Ca2+-ATPase (hSERCA-2a), expressed by Saccharomyces cerevisiae. (en)
Title
  • Avidin Functionalized Maghernite Nanoparticles and Their Application for Recombinant Human Biotinyl-SERCA Purification
  • Avidin Functionalized Maghernite Nanoparticles and Their Application for Recombinant Human Biotinyl-SERCA Purification (en)
skos:prefLabel
  • Avidin Functionalized Maghernite Nanoparticles and Their Application for Recombinant Human Biotinyl-SERCA Purification
  • Avidin Functionalized Maghernite Nanoparticles and Their Application for Recombinant Human Biotinyl-SERCA Purification (en)
skos:notation
  • RIV/61989592:15310/12:33141654!RIV13-MSM-15310___
http://linked.open...avai/predkladatel
http://linked.open...avai/riv/aktivita
http://linked.open...avai/riv/aktivity
  • P(ED2.1.00/03.0058), P(KAN115600801), P(KAN200380801)
http://linked.open...iv/cisloPeriodika
  • 43
http://linked.open...vai/riv/dodaniDat
http://linked.open...aciTvurceVysledku
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http://linked.open...iv/duvernostUdaju
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  • 124322
http://linked.open...ai/riv/idVysledku
  • RIV/61989592:15310/12:33141654
http://linked.open...riv/jazykVysledku
http://linked.open.../riv/klicovaSlova
  • expression; adsorption; oxides; spectra; surfaces; iron; binding; membrane-proteins; polyacrylamide gels; magnetic nanoparticles (en)
http://linked.open.../riv/klicoveSlovo
http://linked.open...odStatuVydavatele
  • US - Spojené státy americké
http://linked.open...ontrolniKodProRIV
  • [38C7790B8EA3]
http://linked.open...i/riv/nazevZdroje
  • Langmuir: the A C S journal of surfaces and colloids
http://linked.open...in/vavai/riv/obor
http://linked.open...ichTvurcuVysledku
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http://linked.open...vavai/riv/projekt
http://linked.open...UplatneniVysledku
http://linked.open...v/svazekPeriodika
  • 28
http://linked.open...iv/tvurceVysledku
  • Zbořil, Radek
  • Baratella, Davide
  • Bertipaglia, Ilenia
  • Faralli, Adele
  • Giannetti, Sara
  • Magro, Massimiliano
  • Salviulo, Gabriella
  • Vianello, Fabio
http://linked.open...ain/vavai/riv/wos
  • 000310484300032
issn
  • 0743-7463
number of pages
http://bibframe.org/vocab/doi
  • 10.1021/la303148u
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  • 15310
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