Highly efficient antibody immobilization with multimeric protein Gs coupled magnetic silica nanoparticles

J. H. Lee; H. K. Choi; J. H. Chang

doi:10.1117/12.893366

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12 September 2011 Highly efficient antibody immobilization with multimeric protein Gs coupled magnetic silica nanoparticles

J. H. Lee, H. K. Choi, J. H. Chang

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Proceedings Volume 8099, Biosensing and Nanomedicine IV; 80990L (2011) https://doi.org/10.1117/12.893366
Event: SPIE NanoScience + Engineering, 2011, San Diego, California, United States

Abstract

This work reports the immobilization of monomeric, dimeric and trimer protein Gs onto silica magnetic nanoparticles for self-oriented antibody immobilization. To achieve this, we initially prepared the silica-coated magnetic nanoparticle having about 170 nm diameters. The surface of the silica coated magnetic nanoparticles was modified with 3- aminopropyl-trimethoxysilane (APTMS) to chemically link to multimeric protein Gs. The conjugation of amino groups on the SiO2-MNPs to cysteine tagged in multimeric protein Gs was performed using a sulfo-SMCC coupling procedure. The binding efficiencies of monomer, dimer and trimer were 77 %, 67 % and 55 % respectively. However, the efficiencies of antibody immobilization were 70 %, 83 % and 95 % for monomeric, dimeric and trimeric protein G, respectively. To prove the enhancement of accessibility by using multimeric protein G, FITC labeled goat-anti-mouse IgG was treated to mouse IgG immobilized magnetic silica nanoparticles through multimeric protein G. FITC labeled goat anti-mouse IgGs were more easily bound to mouse IgG immobilized by trimeric protein G than others. Finally protein G bound silica magnetic nanoparticles were utilized to develop highly sensitive immunoassay to detect hepatitis B antigen.

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J. H. Lee, H. K. Choi, and J. H. Chang "Highly efficient antibody immobilization with multimeric protein Gs coupled magnetic silica nanoparticles", Proc. SPIE 8099, Biosensing and Nanomedicine IV, 80990L (12 September 2011); https://doi.org/10.1117/12.893366

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