title: Efficient lipid delivery to hybridoma culture by use of cyclodextrin in a novel granulated dry-form medium technology.
abstract: Lipids are critical nutrients for high density eukaryotic cell cultivation, but inclusion of lipid components into dry-form media has been technically challenging. Lipid supplements are usually supplied for separate addition after powder reconstitution and filtration, which increases manipulation and risk in a biopharmaceutical manufacturing facility. Advanced Granulation Technology (AGT) is a novel dry-form media format designed for large-scale biopharmaceutical manufacture. All components of a complex formulation are homogeneously incorporated into a single granulated medium that simply requires water addition to yield a complete reconstituted 1X medium. We investigated whether cyclodextrin technology could be combined with the AGT process to deliver usable lipid in a dry medium format. The test lipids were cholesterol and several fatty acids supplied either as an aseptic supplement to liquid media or as part of a complete AGT formulation. The test system utilized a cholesterol auxotroph, NS0, and a protein-free basal formulation (CD Hybridoma Medium), with or without supplemental lipid. Post-filtration recovery of cholesterol from AGT-processed materials compared favorably with media supplemented with liquid lipid concentrates. Cell growth and viability and expression of recombinant protein were equivalent in all test media.
authors.affiliation: Cell Culture Research and Development, GIBCO/Invitrogen Corporation, 3175 Staley Road, Grand Island, New York 14072, USA.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Walowitz
authors.forename: Richard M
authors.initials: RM
authors.lastname: Fike
authors.forename: David W
authors.initials: DW
authors.lastname: Jayme
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D003505:Cyclodextrins / Q000494:pharmacology*; D006825:Hybridomas / Q000166:cytology / Q000187:drug effects* / Q000502:physiology*; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000235:genetics; D050356:Lipid Metabolism; D011208:Powders; D011786:Quality Control; D011994:Recombinant Proteins / Q000096:biosynthesis; D012996:Solutions / Q000494:pharmacology
chemical_list: D017077:Culture Media, Conditioned; D003505:Cyclodextrins; D007074:Immunoglobulin G; D011208:Powders; D011994:Recombinant Proteins; D012996:Solutions