Thermosensitive and Salt-Sensitive Molecularly Imprinted Hydrogel for Bovine Serum Albumin

Abstract

A novel stimuli-responsive protein imprinted polymer for selective recognition of bovine serum albumin is presented. N-[3-(Dimethylamino)propyl]-methacrylamide, which is positively charged in neutral solution and is able to self-assemble onto the template protein through electrostatic interaction, was chosen as the functional monomer. Polymerization was carried out in the presence of N-isopropylacrylamide as an assistant monomer, which resulted in a stimuli-responsive protein imprinted polymer. The template proteins were easily removed by treatment with 500 mmol L(-1) NaCl solution. The influences of the external stimuli, such as temperature and ionic strength, on the polymer affinity were investigated, and a clear conformational memory was observed. The association constant ( Ka) and binding capacity ( Qmax) for the specific interaction between the protein imprinted polymer and the template protein were determined by Scatchard plots and found to be 9.6 x 10(4) L mol(-1) and 4.7 micromol g(-1), respectively. Several proteins different in molecular weight and isoelectric point were employed as reference, and it was shown that the charge effect and the shape memory effect were the major factors affecting the imprint formation and template recognition. Finally, this imprinted polymer was used to purify the bovine serum albumin from the protein mixture and real sample, which demonstrated its high selectivity.