Detection of liver cancer cells by using ELISA and coupling of anti-glypican 3 antibody and magnetite nanoparticles

초록

A method for detecting glypican 3 (GPC3) liver cancer cells by coupling of anti-glypican 3 antibody (anti-GPC3) and magnetite nanoparticles (NPs) was investigated to detect GPC3 by enzyme-linked immunosorbent assay (ELISA) in this study. Magnetite NPs with the average size of 11 nm were synthesized by using co-precipitation method of Fe2+ and Fe3+ in NH<inf>3</inf>·H<inf>2</inf>O solution. First, silica was coated on the magnetite NPs using Stöber method to obtain Fe<inf>3</inf>O<inf>4</inf>/SiO<inf>2</inf> core-shell structures and then 3-aminopropyltriethoxysilane (APTES) was treated on the Fe<inf>3</inf>O<inf>4</inf>/SiO<inf>2</inf> by silanization reaction to achieve Fe<inf>3</inf>O<inf>4</inf>/SiO<inf>2</inf>/APTES nanostructures. After modified by APTES, the nanostructures were activated by glutaraldehyde (GA) to obtain functional groups on the nanostructures surface to bind with anti-GPC3 by covalent immobilization. The UV–vis spectroscopy was carried out to investigate the binding of anti-GPC3 to the NPs and binding efficiency (88.35%) was estimated by the Bradford method. The NPs bound anti-GPC3 (NPs/anti-GPC3) can detect GPC3 by using ELISA at low concentration (0.16 ng/ml). © 2015 The Korean Society of Mineral and Energy Resources Engineers (KSMER).

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