In many microorganisms, flux limitations in oxidative metabolism lead to the formation of overflow metabolites even under fully aerobic conditions. This can be avoided if the specific growth rate is controlled at a low enough value. This is usually accomplished by controlling the substrate feeding profile in a fed-batch process. The present work proposes a control concept which is based on the on-line detection of metabolic state by on-line calculation of mass and elemental balances. The advantages of this method are: 1) the check of measurement consistency based on all of the available measurements, 2) the minimum requirement of a priori knowledge of metabolism, and 3) the exclusive use of simple and established on-line techniques which do not require direct measurement of the metabolite in question. The control concept has been linked to a simple adaptive controller and applied to fed-batch cultures of S. cerevisiae and E. coli, organisms which express different overflow metabolites, ethanol and acetic acid, respectively. Oxidative and oxidoreductive states of S. cerevisiae and E. coli cultures were detected with high precision. As demonstrated by the formation of acetic acid in E. coli cultures, metabolic states could be correctly distinguished for systems for which traditional methods, such as respiratory quotient (RQ), are insensitive. Hence, it could be shown that the control concept allowed avoidance of overflow metabolite formation and operation at maximum oxidative biomass productivity and oxidative conversion of substrate into biomass. Based on mass and elemental balances, the proposed method additionally provides a richness of additional information, such as yield coefficients and estimation of concentrations and specific conversion rates. These data certainly help the operator to additionally evaluate the state of the process on-line.

Microencapsulation of cells in a polymer membrane [e.g., poly(hydroxyethyl methacrylate-co-methyl methacrylate) (HEMA-MMA)] has been proposed as a vehicle for the delivery of therapeutic biomolecules, but cells (especially xenogeneic cells) survive only for short times, limiting the utility of this approach. Murine interleukin-10 (mIL-10) has been shown to downregulate the xenogeneic immune response, and we tested the hypothesis that mIL-10 produced by microencapsulated Chinese hamster ovary (CHO) cells would modulate the transplant-site environment leading to prolonged cell function in a xenogeneic model without other immunomodulatory agents. Prior to encapsulation, CHO cells were genetically engineered to express mIL-10 and a firefly bioluminescence protein, luciferase, which allowed for noninvasive tracking of transplanted cells in vivo with the Xenogen IVIS Imaging System. This nondestructive imaging system was sufficiently sensitive to detect photon signal emitted by a single capsule containing around 800 luciferase-transduced CHO (CHO(LUC)) cells in vitro, and to track changes in luciferase expression in vivo over time. Effective modulation of the transplantation-site environment with mIL-10 secreted from capsules was evident by greater luciferase expression at 10 and 21 days after transplantation relative to encapsulated luciferase-transfected cells that did not produce mIL-10. Longer duration effects require further investigation to extend this proof-of-concept study.