Affinity Based Nano-Magnetic Particles for Purification of Recombinant Proteins in Form of Inclusion Body

BACKGROUND: Protein purification is the most complicated issue in the downstream processes of recombinant protein production; therefore, improved selective purification methods are important. Affinity-based protein purification method using His-tag and Ni-NTA resins is one of the most common strategies. MNPs can be used as a beneficial alternative for Ni-NTA resins. However, there is no data on the capability of MNPs for protein purification from inclusion bodies; this issue is studied here. METHODS: Recombinant His-tagged proteins of EGFP-His and SK-His were expressed in E. coli BL-21 (DE3) in soluble and inclusion body forms, respectively. MNPs including Fe(3)O(4) magnetic core, SiO(2) shell, and Ni(2+) on the surface were synthesized by sol-gel and hydrothermal reactions and then characterized by XRD, VSM, and SEM imaging. Both synthesized Fe(3)O(4)@NiSiO(3) and Fe(3)O(4)@Ni(x)SiO(y )MNPs were employed to purify EGEP-His and SK-His under native and denaturing conditions, respectively. The quantity and purity of purified proteins were analyzed by micro-Bradford assay and SDS-PAGE, respectively. RESULTS: Both synthesized MNPs were spherical and well-dispersed with the size ranging from 290 to 415 nm. Synthesized MNPs contained Fe(3)O(4,) SiO(2) shell, and Ni(2+) on their structures with suitable magnetization properties. Using Fe(3)O(4)@NiSiO(3 )and Fe(3)O(4)@Ni(x)SiO(y )yielded 192 and 188 µg/mg of SK-His, as compared to 207 and 195 µg/mg of EGFP-His, respectively. CONCLUSION: MNPs containing magnetic Fe(3)O(4 )core, SiO(2) shell, and Ni(2+)on their surface are versatile alternatives for Ni-NTA resins in protein purification for proteins expressed in both soluble and inclusion body forms.