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

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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 Förster 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: Fluorometric assays; Magnetic submicron particles; N-biotinyl-N′-(1-naphthyl)-ethylenediamine; Streptavidin

Document Type: Research Article

DOI: http://dx.doi.org/10.1366/12-06963

Affiliations: Unit for Analytical Probes and Protein Biotechnology, Key Laboratory of Clinical Laboratory Diagnostics of the Education Ministry, College of Laboratory Medicine, Chongqing Medical University, Chongiqng 400016, China

Publication date: June 1, 2013

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