Mr. Phelps et al., TECHNOLOGY FOR REGENERABLE BIOSENSOR PROBES BASED ON ENZYME CELLULOSE-BINDING DOMAIN CONJUGATES, Biotechnology progress, 10(4), 1994, pp. 433-440
The application of enzyme-based biosensors for on-line bioprocess moni
toring and control has been slowed by problems relating to the in situ
sterilizability of the probe and the stability of the enzyme componen
t. A novel technology with the potential to address both of these diff
iculties is presented here, The approach is based on the reversible im
mobilization of enzymes conjugated with the cellulose binding domain (
CBD) of cellulases from Cellulomonas fimi. A regenerable biosensor ele
ctrode can be configured with a cellulose matrix onto which the enzyme
-CBD conjugate can be repeatedly loaded (bound by the CBD) and subsequ
ently eluted by perfusing the cellulose matrix with the appropriate so
lution. Glucose oxidase (GOx) conjugated to CBD with glutaraldehyde wa
s used in an experimental glucose biosensor to demonstrate the feasibi
lity of multiple cycles of loading and elution of the conjugate. Micha
elis-Menten enzyme kinetics provided an empirical model for the calibr
ation of the experimental biosensor. The development of a computer-con
trolled prototype glucose biosensor and a fermentation monitoring syst
em based on this approach is discussed.