Figure 6.

General method for synthesis of molecularly imprinted magnetic Fe₃O₄@MIP NPs using an immobilized template protein. In the protocol, the surface of Fe₃O₄ NPs was transformed to a silica shell using tetraethoxysilane (TEOS) to form Fe₃O₄@SiO₂. Then, the surface of Fe₃O₄@SiO₂ was reacted with 3-aminopropyltriethoxysilane (APTES) to introduce amino groups on the surface which were reacted with glutaraldehyde (GA) in the following step to form aldehyde modified Fe₃O₄@SiO₂ NPs. In the next step, template protein was covalently immobilized on the aldehyde modified Fe₃O₄@SiO₂ NPs. In the last step, siloxane co-polymerization by APTES and octyltrimethoxysilane resulted in a polymeric network around the template protein on the surface of the Fe₃O₄@SiO₂ – protein complex. Then, the template was removed from the surface and imprinted cavities which were complementary to the template in size, shape and functional groups were obtained (reproduced from [60] with permission).