Selective capturing and purification of C-reactive protein (CRP) from complex biological fluids plays a pivotal role in studying biological activities of CRP in various diseases. However, obvious nonspecific adsorption of proteins was observed on current affinity sorbents, and thus additional purification steps are often required, which could compromise the recovery of the target protein and/or introduce new impurities. In this study, inspired by the highly specific interaction between CRP and the cell membrane, an excellent anti-biofouling compound 2-(methacryloyloxy)ethyl phosphorylcholine and a highly hydrophilic crosslinker N, N'-methylenebisacrylamide were employed to fabricate a novel cell membrane biomimetic polymer for selective capture of CRP in the presence of calcium ions. Based on the polymer described above, a facile enrichment approach was established after systematic optimization of the washing and elution conditions. With its favorable properties, such as good porosity, weak electrostatic interaction, high hydrophilicity, and biocompatibility, the novel biomimetic polymer exhibits good specificity, selectivity, recovery (near 100%), purity (95%), and a lower nonspecific protein adsorption for CRP in comparison with commercial immobilized p-aminophenyl phosphoryl choline gel and other purification materials. Furthermore, the structural integrity and functionality of CRP in the elution fraction were well preserved and confirmed by circular dichroism spectroscopy, fluorescence spectroscopy, and immunoturbidimetric assay. Finally, the biomimetic polymer was successfully applied to the selective enrichment of CRP from sera of patients with inflammation and rats. The proposed novel enrichment approach based on the versatile biomimetic polymer can be used for effective CRP purification, which will benefit the in-depth study of its biological roles.
Keywords: C-reactive protein; biological fluids; cell membrane biomimetic polymer; selective enrichment; zwitterionic phosphorylcholine.