Design of enzyme immobilized zwitterionic copolymer nanogels and its size effect on electrochemical reaction

For enzyme-based electrochemical devices, an improvement in electron transfer between the enzyme and electrode is important. Thus, we developed a nano-scaled hydrogel that includes an electron mediator and enzyme to realize nano-sized effects that enhance the functions. Three different chain lengths (short, medium, and long) of copolymers composed of 2-methacryloyloxyethyl phosphorylcholine (MPC) and methacrylic acid N-hydroxysuccinimide ester (MNHS; poly(MPC-co-MNHS), PMS) were synthesized via reversible addition-fragmentation chain transfer (RAFT) polymerization. The PMS nanogels can bind to the amino ferrocene (AFc) of the electron mediator and glucose oxidase (GOD) as a catalyst. The mono-dispersive PMS nanogels approximately 200-250 nm in size bound with AFc were prepared with different polymer chain lengths and amounts of AFc (PMMFcX_Y%, X= 'degree of polymerization, 50, 75, 100' and Y= 'AFc feeding ratio against the amount of NHS group in the polymer chain, 50 %, 100 %'). The size of PMMFcX_Y% could be controlled by changing degree of polymerization or AFc feeding ratio. After the modification of GOD to PMMFcX_Y%, their size increased slightly from the original size (ca. 200-250 nm) to approximately 250-300 nm. The catalytic activity of nanogel in dispersed system was higher than that of microgel, indicating that nanogels could improve glucose transport in hydrogel layer. Compared to the catalytic reaction of the PMMFc 75_50 %-GOD nanogel-modified electrodes with that of microgel modified electrode, the current response was improved by decreasing the nanogel size, as evaluated by electrochemical measurements. These results revealed that the smaller nanogels could improve both glucose transport and electron transfer via mediator by smaller size, resulting higher efficiency of enzyme immobilized electrode.