title: Monolithic molecularly imprinted cryogel for lysozyme recognition.
abstract: The application of molecularly imprinted polymers in the selective adsorption of macromolecules such as proteins by monolithic protein-imprinted columns requires a macroporous structure, which can be provided by cryogelation at low temperature in which the formation of ice crystals gives a porous structure to the molecularly imprinted polymer. In this study, we applied this technique to synthesize lysozyme-imprinted polyacrylamide cryogels containing 8% w/v of total monomers and 0.3% w/v of lysozyme. The synthesized cryogel was sponge-like and elastic with very fast swelling and reshaping properties, showing a swelling ratio of 24.5 ± 3 and gel fraction yield of about 72%. It showed an imprinting effect of 1.58 and a separation factor of 1.37 for cytochrome c as the competing protein. Adsorption studies on the cryogel revealed that it follows the Langmuir isotherm, with a maximum theoretical adsorption capacity of 36.3 mg lysozyme per gram of cryogel. Additionally, it was shown that a salt-free rebinding solution at low flow rate and pH = 7.0 is favorable for lysozyme rebinding. This kind of monolithic column promises a wide range of application in separation of various biomolecules due to its preparation simplicity, good rebinding characteristics, and macroporosity.
authors.affiliation: Biotechnology Research Laboratory, School of Chemical Engineering, Iran University of Science and Technology (IUST), Tehran, Iran.
authors.forename: Mohammadmahdi
authors.initials: M
authors.lastname: Rabieizadeh
authors.forename: Seyed Mohammadreza
authors.initials: SM
authors.lastname: Kashefimofrad
authors.forename: Fereshteh
authors.initials: F
authors.lastname: Naeimpoor
mesh_terms: D000327:Adsorption; D059985:Cryogels / Q000737:chemistry*; D008855:Microscopy, Electron, Scanning; D054802:Molecular Imprinting; D009113:Muramidase / Q000737:chemistry*; D013816:Thermodynamics
chemical_list: D059985:Cryogels; D009113:Muramidase
keywords: Cryogel; Lysozyme; Molecular imprinting; Monolithic columns