Aptamer functionalized hydrophilic polymer monolith with gold nanoparticles modification for the sensitive detection of human α-thrombin.

Low abundant proteins of body fluids participate nearly all physiological processes and indicate various kinds of diseases. The development of specific enrichment techniques is the key to identify and quantify the low abundant proteins. Herein, a novel kind of aptamer functionalized hydrophilic polymer monolith was developed for the specific enrichment and detection of human α-thrombin from the human plasma. Human α-thrombin aptamer, with thiol group modified at the 5' terminal, was immobilized on the gold nanoparticles (AuNPs) modified poly(glycidyl methacrylate-co-poly(ethylene glycol) diacrylate) monolithic column, with the binding capacity of 277.1μmol/L. Due to the hydrophilic poly(ethylene glycol) diacrylate) as the cross-linking monomer, the detection recovery of the aptamer-functionalized hydrophilic polymer monolithic column could reach to 92.6±5.2% (n=3) and the dynamic range could reach 0.5-300ng/μL (S/N>10) with on-line UV detection. Meanwhile, the column could run over 100 times, because the poly(glycidyl methacrylate-co-poly(ethylene glycol) diacrylate) stability structure and the AuNPs improved the stability of the matrix material. Furthermore, this column could even capture the target α-thrombin, which was spiked in 1000 folds of original human plasma. All these results demonstrated the great potential of the prepared aptamer functionalized hydrophilic polymer monolith for the recognition of the trace proteins in the biological samples.