Molecularly imprinted polymers for the recognition of sodium dodecyl sulfate denatured creatine kinase

Micro-contact imprinting methodology was used to prepare the molecule recognition film for denatured creatine kinase-MM (CK-MM), which was denatured by sodium dodecyl sulfate. Interpretation of thermo-calorimetry measurements suggested the use of poly(ethylene glycol) 400 dimethacrylate (PEG400DMA) as a crosslinking molecule. The selected functional monomer, methyl methacrylate (MMA), produced the highest imprinting factor from a panel of five candidates. The molecularly imprinted polymers (MIPs) formed with 5% MMA, 95% PEG400DMA and denatured OK-MM showed excellent imprint recognition, with the imprinting factor of 8.66. The dissociation constant calculated from Scatchard plot was 3.25 x 10(-8) M. MIPs had little affinity with non-template proteins, such as native CK-MM, a native and denatured form of human serum albumin (HSA) and immunoglobulin G (IgG). The competitive study in a two-protein environment showed that the selectivity of MIP was 96.8% and 98.7% for denatured CK-MM/denatured HSA and denatured CK-MM/denatured IgG, respectively. The results suggested that MIPs prepared in this work recognize not only the protein sequences but also the protein secondary structure. (C) 2011 Taiwan Institute of Chemical Engineers. Published by Elsevier B.V. All rights reserved,