title: Disposable polymeric cryogel bioreactor matrix for therapeutic protein production.
abstract: Low cost and high efficiency make disposable bioreactors feasible for small-scale therapeutic development and initial clinical trials. We have developed a cryogel-based disposable bioreactor matrix, which has been used for production of protein therapeutics such as urokinase and monoclonal antibodies (mAbs). The protocol discusses the application of a cryogel bioreactor for mAb production. Cryogels composed of either polyacrylamide (PAAm) coupled to gelatin or semi-interpenetrating PAAm-chitosan are synthesized by free-radical polymerization at -12 °C. Hybridoma cells are immobilized over the cryogel bioreactor and incubated for 48 h. Medium is circulated thereafter at 0.2 ml min(-1) and bioreactors can be run continuously for 60 d. The cryogel-based packed-bed bioreactor can be formulated as a monolith or as beads; it also has an efficiency four times what can be obtained using a tissue-culture flask, a high surface-to-volume ratio and effective nutrient transport. After incubation, the bioreactor setup will take about 60 min using a pre-prepared sterilized cryogel.
authors.affiliation: Department of Biological Sciences and Bioengineering, Indian Institute of Technology Kanpur, Kanpur, India.
authors.forename: Era
authors.initials: E
authors.lastname: Jain
authors.forename: Ashok
authors.initials: A
authors.lastname: Kumar
mesh_terms: D000180:Acrylic Resins / Q000138:chemical synthesis; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D019149:Bioreactors; D018929:Cell Culture Techniques; D002460:Cell Line; D059985:Cryogels; D004209:Disposable Equipment; D006801:Humans; D006825:Hybridomas; D014568:Urokinase-Type Plasminogen Activator / Q000096:biosynthesis; D000099166:Polyacrylamides
chemical_list: D000180:Acrylic Resins; D000911:Antibodies, Monoclonal; D059985:Cryogels; D014568:Urokinase-Type Plasminogen Activator; D000099166:Polyacrylamides
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