Immobilization of hybridoma cells in chitosan alginate beads

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Abstract
A new method for immobilizing hybridoma cells using chitosan‐stabilized calcium alginate beads was developed. The ionotropic gelation of chitosan and calcium with alginate resulted in the formation of highly cross‐linked, porous beads that were mechanically and chemically stable in phosphate buffered medium. Hybridomas entrapped in these beads were cultured semi‐continuously using periodic medium exchange. Viable population densities in the order of 5 × I07 cells/mL were attained within the beads and up to two‐fold increases in volumetric monoclonal antibody (MAb) productivity over batch suspension cultures were observed. Oxygen mass transfer limitations within the chitosan‐alginate beads were evaluated by considering biokinetics and diffusive transport. Model equations were developed and used to evaluate the effect of bead diameter on the contained cells. The predictions were consistent with experimental observations of maximum viable population densities attained in beads of various size.