Magnetic functionalized
chitosan-based nanocarrier proffered to purify protein and enzyme has
incepted a striking place in separation and purification predominantly
for its convenient and rapid performance. Here, the surface modification
of chitosan biopolymer (CS) was employed by terephthaloyl chloride, dopamine (DA), and NiCl2 (Ni2+), followed by an in-situ magnetization by Fe3O4 nanoparticles (NPs). This magnetic nanocomposite was utilized for alkaline phosphatase
separation from bovine milk. Based on the FE-SEM images, the particle
size narrow range between 55.4 nm and 72.1 nm was measured, and the
spherical NPs represented a mean diameter of 66.9 nm. Moreover, the
reduction in the magnetization saturation (MS) of synthesized CS-DA/Ni2+/Fe3O4 nanocomposite (3.18 emu/g) compared to Fe3O4 MNPs’ MS (44.2 emu/g) is correlated to the addition of non-magnetic shells. The activity measurements of the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and alkaline phosphatase
(ALPs) were administrated for purification process investigation.
Considering the purification yield of the extracted protein
concentration by mentioned nanocomposite, the most efficient ALPs
protein purification was about 67% after 30 min incubation at 30 °C
through a cost-efficient and reproducible approach. Due to the affinity
purification, the Ni2+ affinity ligand
immobilized on the nanocarrier presented high stability. After five
cycles, a very low adsorption capacity reduction was detected, and 94%
of the original binding capacity of the magnetic nanocomposite was
retained, indicating high reusability of the nanocomposite and the
specified affinity ligand selection toward ALPs.