Facile Characterization of the Immobilization of Streptavidin on Magnetic Submicron Particles with a Fluorescent Probe of Streptavidin

Abstract

To characterize streptavidin immobilization on magnetic submicron particles (MSPs), residual streptavidin after magnetic removal of immobilized streptavidin was quantified with N-biotinyl-N′-(1-naphthyl)-ethylenediamine (BNEDA) based on Forster resonance energy transfer. Residual BNEDA after magnetic removal of bound BNEDA was measured by its own fluorescence. Streptavidin was immobilized at about 12 mg per gram of MSPs and easily retained over 50% of its original activity. These assays facilitated optimized streptavidin immobilization on MSPs.

Keywords

Magnetic submicron particles Fluorometric assays Streptavidin

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References

Magnani

Galluzzi

Bruce

I.J.

. “The Use of Magnetic Nanoparticles in the Development of New Molecular Detection Systems”. J. Nanosci. Nanotechnol. 2006. 6(8): 2302–2311.

Horak

Babic

Mackova

Benes

M.J.

. “Preparation and Properties of Magnetic Nano- and Microsized Particles for Biological and Environmental Separations”. J. Sep. Sci. 2007. 30(11): 1751–1772.

Jung

J.H.

Lee

J.H.

Shinkai

. “Functionalized Magnetic Nanoparticles as Chemosensors and Adsorbents for Toxic Metal ions in Environmental and Biological Fields”. Chem. Soc. Rev. 2011. 40(9): 4464–4474.

Liao

Xie

Y.L.

Yang

X.L.

Deng

Chen

Y.W.

Xie

G.M.

Zhu

Liu

B.Z.

Yuan

H.D.

Liao

Zhao

Y.S.

M.A.

. “Homogeneous Noncompetitive Assay of Protein via Forster-Resonance-Energy-Transfer with Tryptophan Residue(s) as Intrinsic Donor(s) and Fluorescent Ligand as Acceptor”. Biosens. Bioelectron. 2009. 25(1): 112–117.

Xie

Y.L.

Yang

X.L.

Zhao

Y.S.

Zhang

Xie

G.M.

Zheng

Yuan

H. D.

Liao

. “Homogeneous Competitive Assay of Ligand Affinities Based on Quenching Fluorescence of Tyrosine/Tryptophan Residues in a Protein via Forster-Resonance-Energy-Transfer”. Spectrochim. Acta A 2010. 77(4): 869–876.

Yang

X.L.

Xie

Y.L.

Zhao

Liao

Chen

Y.W.

Long

G.B.

Zhang

Yuan

Y.H.

Zhu

Yuan

H.D.

Liao

. “Estimation of Affinities of Ligands in Mixtures via Magnetic Recovery Of Target-Ligand Complexes and Chromatographic Analyses: Chemometrics and an Experimental Model”. BMC Biotechnol. 2011. 11: 44. doi: 10.1186/1472-6750-11-44.

Yang

X.L.

Zhao

X.Y.

Liao

Xie

Y.L.

Zhu

Long

G.B.

Yuan

Y.H.

Liao

. “Method to Screen Aromatic Ligands in Mixtures for Quantitative Affinities to Target Using Magnetic Separation of Bound Ligands Along with HPLC and UV Photometry Detection”. Microchim. Acta 2012. 176(1-2): 243–249.

Gupta

A.K.

Gupta

. “Synthesis and Surface Engineering of Iron Oxide Nanoparticles for Biomedical Applications”. Biomaterials 2005. 26(18): 3995–4021.

Osaka

Matsunaga

Nakanishi

Arakaki

Niwa

Iida

. “Synthesis of Magnetic Nanoparticles and Their Application to Bioassays”. Anal. Bioanal. Chem. 2006. 384(3): 593–600.

10.

Frey

N.A.

Peng

Cheng

Sun

S.H.

. “Magnetic Nanoparticles: Synthesis, Functionalization, and Applications in Bioimaging and Magnetic Energy Storage”. Chem. Soc. Rev. 2009. 38(9): 2532–2542.

11.

Sandhu

Handa

Abe

. “Synthesis and Applications of Magnetic Nanoparticles for Biorecognition and Point of Care Medical Diagnostics”. Nanotechnology 2010. 21(24): 442001.

12.

Selvan

S.T.

Tan

T.T.

D.K.

Jana

N.R.

. “Functional and Multifunctional Nanoparticles for Bioimaging and Biosensing”. Langmuir 2010. 26(14): 11631–11641.

13.

Behrens

. “Preparation of Functional Magnetic Nanocomposites and Hybrid Materials: Recent Progress And Future Directions”. Nanoscale 2011. 3(3): 877–892.

14.

Wei

Insin

Lee

Han

H.S.

Cordero

J.M.

Liu

Bawendi

M.G.

. “Compact Zwitterion-Coated Iron Oxide Nanoparticles for Biological Applications”. Nano Lett. 2012. 12(1): 22–25.

15.

Stetsenko

D.A.

Gait

M.J.

. “A Convenient Solid-Phase Method for Synthesis of 3′-Conjugates of Oligonucleotides”. Bioconjug. Chem. 2001. 12(4): 576–586.

16.

Henegariu

Bray-Ward

D.C.

. “Custom Fluorescent-Nucleotide Synthesis as an Alternative Method for Nucleic Acid Labeling”. Nat. Biotechnol. 2000. 8(3): 345–348.

17.

De Palma

Trekker

Peeters

Van Bael

M.J.

Bonroy

Wirix-Speetjens

Reekmans

Laureyn

Borghs

Maes

. “Surface Modification of Gamma-Fe₂O₃@SiO₂ Magnetic Nanoparticles for the Controlled Interaction with Biomolecules”. J. Nanosci. Nanotechnol. 2007. 7(12): 4626–4641.

18.

Mittal

Bruchez

. “Biotin-4-Fluorescein Based Fluorescence Quenching Assay for Determination of Biotin Binding Capacity of Streptavidin Conjugated Quantum Dots”. Bioconjug. Chem. 2011. 22(3): 362–368.