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  • Author or Editor: R. Kemp x
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Abstract  

In the photomicrocalorimetric module designed by Johansson and Wadsö for a commercial Thermometric TAM heat conduction batch microcalorimeter, the incident light from an external xenon lamp was divided by a beam splitter and directed to the two vessels of the differential system by light guides ideally to give zero heat flow. In practice this proved difficult and so to improve the balance between the vessels in terms of the incident light heat output as well as potentially to give more versatility regarding the choice of wavelengths, the xenon lamp-based system was replaced in the first stage by a pair of cold white LEDs embedded directly in the test and reference vessels. The LEDs had independent electrical circuits to achieve the balance by manual adjustment. As a second stage, the test vessel was equipped with PTFE tubing for changing the liquid phase in it while it was in the middle thermal equilibrium position. This improved the reproducibility of the results.

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Abstract  

After a formal explanation of Mayer's enthalpy balance method as applied to biological reaction rates, the history of its application is traced from Rubner's dog to accounting for the energy of muscle contraction. The introduction of microcalorimetry allowed the method generally to be used for cells in vitro and now particular emphasis can be paid to the growth of cells for the production of therapeutically-important heterologous proteins. In these systems, enthalpy balance studies contribute to defining catabolic processes, designing media, understanding the mechanisms of growth and controlling cultures using heat flux as an on-line sensor of metabolic activity.

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Abstract  

It is claimed, though not without dispute, that genetically engineered mammalian cells grow more slowly than their progenitor cells because the recombinant gene system causes a metabolic burden. This was found to be the case for CHO cells transfected with expression vectors forcytochrome b5. The slower growth was associated with lower metabolic activity measured by heat flux and mitochondrial activity (rhodamine 123 fluorescence). The calorimetric-respirometric ratio was similar for all cell types, implying that the greater fluxes of glucose and glutamine in the recombinant cells was channelled to biosynthesis. This demand probably restricted the supply of pyruvate to the mitochondria in these cells.

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Abstract  

Microcalorimeters to monitor the heat dissipation of bench-scale animal cell cultures on line and in real time require a continuous circuit between the vessel measuring heat flow rate and the bioreactor. The modifications to the transmission lines and calorimetric heat exchanger were to: (i) reverse the usual upward direction of the cell suspension in the flow vessel to downwards; (ii) install an in situ washing/cleaning facility; (iii) use low diffusivity PEEK material; and (iv) maintain thermal equilibration by water-jacketing the transmission tubing. Chemical calibration showed that there was more than a 20% difference between the physical volume and the effective thermal volume. An appropriate thermodynamic system was defined in order to permit enthalpy balance studies.

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