Dynamic Thermoelectric Glucose sensing with Layer-by-Layer Glucose Oxidase Immobilization

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  • Detecting the loss or generation of stoichiometrically related chemical species with an appropriate electrode. Stability of sensor depends upon the stability of the immobilized enzyme.
  • Coefficient of determination or squared of correlation coefficient.
  • Dynamic Thermoelectric Glucose sensing with Layer-by-Layer Glucose Oxidase Immobilization

    1. 1. Dynamic Thermoelectric Glucose Sensing with Layer-by-Layer Glucose Oxidase Immobilization<br />Siva Mahesh Tangutooru1<br />V.L. Kopparthy, R. Gumma, G.G. Nestorova, E.J. Guilbeau<br />Center for Biomedical Engineering and Rehabilitation Science<br />Louisiana Tech University<br />
    2. 2. Goal<br />To develop a microfluidic glucose calorimeter by immobilizing glucose oxidase using layer-by-layer self-assembly.<br />
    3. 3. Presentation Summary<br />Principle of thermoelectricity<br />Calorimeter design and measurement system<br />Layer-by-layer self-assembly procedure<br />Results<br />Conclusion<br />Future work<br />
    4. 4. Thermoelectric detection of glucose<br />𝐺𝑙𝑢𝑐𝑜𝑠𝑒+𝑂2+𝐻2𝑂𝑔𝑙𝑢𝑐𝑜𝑠𝑒 𝑜𝑥𝑖𝑑𝑎𝑠𝑒𝐺𝑙𝑢𝑐𝑜𝑛𝑖𝑐 𝑎𝑐𝑖𝑑+𝐻2𝑂2+79 𝑘𝐽/𝑚𝑜𝑙<br /> <br />
    5. 5. Outlet<br />Inlet1<br />Device fabrication<br />Inlet2<br />Thermopile<br />Microfluidic Device<br />Microfluidic Calorimeter<br /> PDMS inlet & outlet connectors<br /> Microscope glass slide<br />Kapton® tape<br /> Microscope glass coverslip<br />(a)<br />(a)<br />(b)<br />(c)<br />(d)<br />
    6. 6. Experimental measurement system<br />
    7. 7. Hydrodynamic focusing<br />G. G. Nestorova and E. J. Guilbeau, "Thermoelectric method for sequencing DNA," Lab on a Chip, vol. 11, pp. 1761-1769.<br />
    8. 8. PEI<br />Layer-by-layer self assembly<br />PSS<br />Glucose Oxidase<br />PEI<br />Layer-by-layer electrostatic adsorption mechanism. <br />General adsorption procedure of ployelectrolytes on a substrate. <br />Layer-by-layer assembly of glucose oxidase on the substrate. <br />Structure of polyelectrolytes used for LbL assembly.<br />N+<br />H2<br />SO3- Na+<br />Polyelectrolyte<br />Glucose<br />Oxidase<br />Immobilization procedure for LbL assembly.<br />K. Ariga, Y. Lvov “Self-Assembly of Functional Protein Multilayers: From Flat Films to Microtemplate Encapsulation”, “Biopolymers at Interfaces” Ed. M. Malmsten, M. Dekker Publ., 2003, NY, p.367-391.<br />(a)<br />PSS<br />PEI<br />(c)<br />(b)<br />
    9. 9. Our novel glucose calorimeter<br />Calorimeter<br />Self-generating signal<br />No external power requirement<br />Relatively inexpensive<br />Small in size and light weight<br />High rejection of common mode thermal signals<br />Relatively simple to manufacture<br />Rugged and durable<br />Layer-by-layer Immobilization<br />Multiple layers are physically adsorbed.<br />Easy preparation and high bond strength.<br />
    10. 10. Sensor response for glucose concentration<br />75 mg/dL<br />100 mg/dL<br />Flow rates: 100 µlmin-1 and 25 µlmin-1<br />No. of Immobilized glucose oxidase layers: 2<br />
    11. 11. Effect of number of immobilized layers on sensor response<br />Flow rates: 100 µlmin-1 and 25 µlmin-1<br />
    12. 12. Effect of flow rates on sensor response<br />No. of Immobilized glucose oxidase layers: 2<br />
    13. 13. Conclusion<br />Successfully immobilized multiple layers of glucose oxidase using LbL self-assembly.<br />Multiple layers of glucose oxidase had little effect because oxygen limited the reaction. <br />Increasing the ratio of inlet flow rates improved calorimeter response.<br />Future work<br />This thermoelectric method can be employed to detect the enthalpy produced by enzymatic reactions.<br />glutamate concentration detection in the flow stream by immobilizing glutamate oxidase using layer-by-layer self-assembly.<br />
    14. 14. Thank you<br />Questions ???<br />
    15. 15. M. J. Muehlbauer, E. J. Guilbeau, and B. C. Towe, "Applications and stability of a thermoelectric enzyme sensor," Sensors & Actuators: B. Chemical, vol. 2, pp. 223-232, 1990.<br />

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