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

Abstract

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 NH₃·H₂O solution. First, silica was coated on the magnetite NPs using Stöber method to obtain Fe₃O₄/SiO₂ core-shell structures and then 3-aminopropyltriethoxysilane (APTES) was treated on the Fe₃O₄/SiO₂ by silanization reaction to achieve Fe₃O₄/SiO₂/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).

Keywords::

• detection
• glypican 3
• magnetite nanoparticles
• ELISA

Acknowledgements

The authors acknowledge financial support from Can Tho Department of Science and Technology. The authors would like to thank Nam Khoa Biotek Company; HCM City Institute of Physics and University of Medicine and Pharmacy HCM City for assistive devices.

Disclosure statement

No potential conflict of interest was reported by the authors.