Incorporation of styrene enhances recognition of ribonuclease A by molecularly imprinted polymers.
Ribonuclease A (RNase A) is an RNA-cleaving enzyme characterized by its high conformational stability and strong catalytic activity. This enzyme is ubiquitous in living organisms and is difficult to inactivate. In polymerase chain reaction (PCR) RNase activity is removed by adding inhibitors. Molecularly imprinted polymers (MIPs) with high selectivity, high stability, low cost and facile synthesis could prove useful in extraction of target molecules, such as RNase A, from reaction mixtures. In this investigation, MIPs were synthesized from the monomers styrene and polyethyleneglycol 400 dimethacrylate (PEG400DMA) in several different ratios. Styrene as a functional monomer gave MIPs with a higher affinity for RNase A than other functional monomers tested, according to both enzyme-linked immnuosorbent assay (ELISA) and isothermal titration calorimetry (ITC). The optimum volume ratio of styrene/PEG400DMA was 20/100 at 25 degrees C, and this ratio maximized the rebinding efficiency of RNase A to MIPs. Isothermal titration calorimetry was also used, and could be useful to design the composition of molecularly imprinted polymers for various target molecules.
Hsu
Chung-Yi
CY
Department of Chemical Engineering, National Cheng Kung University, Tainan 701, Taiwan, ROC.
Lin
Hung-Yin
HY
Thomas
James L
JL
Wu
Bo-Tan
BT
Chou
Tse-Chuan
TC
D000327:Adsorption; D001665:Binding Sites; D015374:Biosensing Techniques / Q000379:methods*; D020099:Coated Materials, Biocompatible / Q000737:chemistry*; D008422:Materials Testing; D011092:Polyethylene Glycols / Q000737:chemistry*; D011485:Protein Binding; D012259:Ribonuclease, Pancreatic / Q000737:chemistry*; D020058:Styrene / Q000737:chemistry*; D013499:Surface Properties
D020099:Coated Materials, Biocompatible; D011092:Polyethylene Glycols; D020058:Styrene; D012259:Ribonuclease, Pancreatic