Preparation of C-Terminal Epitope Imprinted Particles Via Reversible Addition-Fragmentation Chain Transfer Polymerization and Zn2+ Chelating Strategy: Selective Recognition of Cytochrome c

The C-terminal epitope imprinted polymers on the silica were prepared by reversible addition-fragmentation chain transfer (RAFT) strategy with C-terminal nonapeptide of cytochrome c (Cyt c) as the template. 4-cyano-4-(phenylcarbonothioylthio) phetanoic acid was immobilized on the silica as a chain transfer agent to regulate the polymerization of the imprinted layer to enhance the recognition performance of the imprinted materials for the peptide and target protein. Additionally, zinc methacrylate was used as a functional monomer to form the imprinted sites through the synergistic effect of Zn2+ chelating, hydrogen bonding and electrostatic attraction, which further improved the recognition between polymers and protein. The molecularly imprinted polymers prepared with RAFT strategy show good recognition for the epitope peptide and Cyt c with imprinting factors of 7.96 and 6.17, respectively, which is higher than those prepared without RAFT strategy. Furthermore, the imprinted polymers have a good binding capacity and imprinting factor for Cyt c in multi-proteins selective recognition and have good reusability. The performance of Cyt c recognition in bovine serum by the epitope imprinted polymers was calculated by HPLC and the results demonstrated the well selectivity and potential application for Cyt c recognition in biological sample.