Magnetic silica nanoparticle-Taq polymerase hybrids for multiple uses in polymerase chain reaction


Ozalp V. C. , Bayramoglu G., Arica M. Y.

RSC ADVANCES, vol.5, no.106, pp.87672-87678, 2015 (Journal Indexed in SCI) identifier identifier

  • Publication Type: Article / Article
  • Volume: 5 Issue: 106
  • Publication Date: 2015
  • Doi Number: 10.1039/c5ra15677g
  • Title of Journal : RSC ADVANCES
  • Page Numbers: pp.87672-87678

Abstract

Enzyme-magnetic nanoparticle hybrids have been employed as a strategy to improve the biocatalytic usability of biological molecules. For this purpose, we synthesized magnetic core-silica shell nanoparticles for direct one-step easy immobilization of Taq polymerase from a crude extract preparation. The hybrid magnetic particles were synthesized by a magnetic co-precipitation method. The magnetic particles were then coated with a 3-(triethoxysilyl)-propylamine (APTES)/tetraethoxysilane (TEOS) mixture, and the pendant -NH2 groups subsequently functionalized with glutaraldehyde for simultaneous immobilization of Taq-polymerase. The magnetic properties of the particles contributed to fast purification to eliminate inhibitory elements present in the crude extract during Taq polymerase isolation. The Taq-silica hybrid material performed with a similar efficiency to the solution-phase enzyme. Additionally, the new hybrid material allowed reuse of the enzyme multiple times. The silica-Taq polymerase hybrid lost 16% of its activity after 6 cycles. Most importantly, the silica microparticle immobilization extended the functional life of Taq polymerase at room temperature by facilitating a fast cleaning procedure.