Biotechnology and Bioprocess Engineering:BBE

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
권호 표지

Immobilization of Proteins on Magnetic Nanoparticles

  • Wang, Tzu-Hsien (Department of Chemical Engineering, National Chung Cheng University) ;
  • Lee, Wen-Chien (Department of Chemical Engineering, National Chung Cheng University)
  • Published : 2003.07.01
Download PDF
⟨ Previous Next ⟩

Abstract

Magnetic nanoparticles prepared from an alkaline solution of divalent and trivalent iron ions could covalently bind protein via the activation of Nethyl-N-(3-dimethylaminopropyl) carbodiimide (EDC). Trypsin and avidin were taken as the model proteins for the formation of protein-nanoparticle conjugates. The immobilized yield of protein increased with molar ratio of EDC/nanoparticie. Higher concentrations of added protein could yield higher immobilized protein densities on the particles. In contrast to EDC, the yields of protein immobilization via the a ctivation of cyanamide were relatively lower. Nanoparticles bound with avidin could attach a single-stranded DNA through the avidin-biotin interaction and hybridize with a DNA probe. The DNA hybridization was confirmed by fluorescence microscopy observations. Immobilized DNA on nanoparticles by this technique may have widespread applicability to the detection of specific nucleic acid sequence and targeting of DNA to particular cells.

Keywords

References

  1. J.Magn.Magn.Mater v.122 Biological and biomedical aspects of magnetic fluid technology Roath,S https://doi.org/10.1016/0304-8853(93)91103-E
  2. Langmuir v.15 Preparation and properties of magnetite and poly-mer magnetite nanoparticles Dresco,P.A;V.S.Zaitsev;R.J.Gambino;B.Chu https://doi.org/10.1021/la980971g
  3. Bioconjugate Chem v.4 An-tibody-magnetite nanoparticles:In vitro characterization of a potential tumor-specific contrast agent for magnetic resonance imaging Tiefenauer,L.X;G.Kuhne;R.Y.Andres https://doi.org/10.1021/bc00023a007
  4. Bioconju-gate Chem v.10 High-effciency intracellular magnetic labeling with novel superparamagnetic-tat peptide conjugates Josephson,L;C.-H.Tung;A.Moore;R.Weissleder https://doi.org/10.1021/bc980125h
  5. Electrophoresis v.19 Surface modification of superparamag-netic nanoparticles(Ferrofluid) studies with particle elec-trophoresis:Application to the specific targeting of cells Sestier C;M.F.Da-Silva;D.Sabolovic;J.Roger;J.N.Pons https://doi.org/10.1002/elps.1150190725
  6. Biotechnol.Tech v.11 Direct binding of protein to magnetic particles Mehta R.V;R.V.Upadhyay;S.W.Charles;C.N.Ramchand https://doi.org/10.1023/A:1018457915380
  7. J.Magn.Magn.Mater v.201 Immobilization of protein and enzymes to fine magnetic particles Koneracka M;P.Kopcansky;M.Antalmk;M.Timko;C.N.Ramchand;D.Lobo;R.V.Mehta;R.V.Upadhyay https://doi.org/10.1016/S0304-8853(99)00005-0
  8. Biotechnol.Lett v.23 Immobilization of yeast alcohol dehydrigenase on magnetic nanoparticles for improving its stability Liao,M.-H;D.-H.Chen https://doi.org/10.1023/A:1012485221802
  9. J.Mol.Catal.B:Enzymatic v.16 Preparation and characterization of YADH-bound magnetic nanoparticles D.-H.Chen;M.-H.Liao https://doi.org/10.1016/S1381-1177(01)00074-1
  10. Nucleic Acids Res v.16 Assessment of methods for covalent binding of nucleic acids to magnetic beads,Dynabeads,and the characteris-tics of the bound nucleic acid in hybridization reactions Lund,V;R.Schmid;D.Rickwood;E.Hornes https://doi.org/10.1093/nar/16.22.10861
  11. Anal.Biochem v.236 Amination of polystyrene mi-crowells:application to the covalent grafting of DNA probes for hybridization assays Zammatteo,N;C.Girardeaux;D.Delforge;J.J.Pireauz,J.Remacle https://doi.org/10.1006/abio.1996.0135