A Portable Visual Sensor by Molecularly Imprinted Hydrogels for HRP Recognition

Background: Molecular imprinting is a technology used to produce artificial receptors that simulate the molecular recognition in the nature and prepare the polymer network structure in the presence of template molecule. Molecularly imprinted visual sensor combines the advantages of specific recognition via molecular imprinting and fast response speed via visualization. The aims of this paper are to prepare a portable visual sensor for Horseradish Peroxidase (HRP) recognition based on molecularly imprinted hydrogel. Methods: At first, HRP-imprinted polyacrylamide hydrogels with 1 mm thickness were obtained by one-step synthesis via radical induced in-situ polymerization of acrylamide using acrylamide (AAm) as the functional monomer, N,N'-Methylenebisacrylamide (MBA) as the crosslink agent and HRP as the template molecule. Results: Compared with nonimprinted hydrogels, the HRP-imprinted hydrogel sensor showed significant color changes in response to the target HRP. This visual sensor was constructed based on 3, 3', 5, 5'- tetramethyl benzidine (TMB) - H2O2 color reaction system by HRP catalyzing to produce color change through digital photography and image analysis (RGB system). The HRP-imprinted hydrogel showed good response in the range of 0.001-0.5 mg/mL and had a significant specific recognition compared to other proteins via selective test. Conclusion: The proposed portable visual sensor could be used for qualitative and semi-quantitative analysis of HRP with high selectivity and reasonable regeneration. The sensor has the advantages of simple operation, low cost, no special equipment, and can be applied to serum sample with less sample consumption and no need of sample preparation. It has wide application prospects in microfluidic devices, biomimetic sensors, flexible biosensor and membrane separation technology.