Preparation of anti-human cardiac troponin I immunomagnetic nanoparticles and biological activity assays

Maghemite nanoparticles (MNPs) were synthesized by chemical coprecipitation and coated with meso-2,3-dimercaptosuccinic acid (HOOC-CH(SH)-CH(SH)-COOH or DMSA). The morphology and properties of the nanoparticles were characterized by TEM, XRD, Zeta Potential Analyzer and VSM. Subsequentially, the anti-human cardiac troponin I (cTnI) immunomagnetic nanoparticles (IMNPs) were prepared by grafting anti-human cTnI antibodies on the surface of DMSA-coated MNPs using the linker of EDC (1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride). The conjugation amount of the antibodies and the activity of IMNPs was evaluated by enzyme linked immunosorbent assay (ELISA) and Western blotting. The results show that the physical and chemical adsorption occurred at the same time, but the former was unstable and apt to desorb, and the maximum conjugation amount of antibody was about 96 μg on the 0.1 mg MNPs by covalent bond. The stability was also investigated, and after 300 days the antibodies on the IMNPs remained the biological activity.