Polydopamine imprinted magnetic nanoparticles as a method to purify and detect class II hydrophobins from heterogeneous mixtures.

Hydrophobins are one of the most active surface active proteins in nature, with an amphiphilic nature and the ability to self-assembly in elastic monolayers, the possible applications in industry are continuously increasing. However, production and purification of these proteins still remains a tedious process. We introduce here the use of polydopamine as imprinter polymer to create specific magnetic nanoparticles for the recognition of Hydrophobin HFBII from Trichoderma reesei. The protein was molecularly imprinted to magnetic nanoparticles to facilitate its specific detection and purification from liquids or carbonated beverages in the presence of other proteins. The resulting magnetic nanoparticles were successfully imprinted adsorbing till 77,4µg of HFBII hydrophobin per miligram of nanoparticles. The adsorption capacity of the imprinted nanoparticles was also tested for specificity using a mixture of five different proteins and peptides. A slight cross interaction was observed when proteins of similar molecular weight to HFBII were used. With larger proteins and peptides the interaction was very low. with other class II Hydrophobins the interaction was very similar as to HFBII.