title: Immunomodulation by LFA3TIP, an LFA-3/IgG1 fusion protein: cell line dependent glycosylation effects on pharmacokinetics and pharmacodynamic markers.
abstract: LFA3TIP, a fusion protein comprised of the first extracellular domain of LFA-3 fused to the hinge, CH2, and CH3 domains of human IgG1 inhibits responses of human and non-human primate T cells in vitro. In seeking to optimize the expression efficiency to prepare large quantities of LFA3TIP for primate studies, the protein was produced in both the CHO (Chinese hamster ovary) and murine NS-0 myeloma cell lines. Although LFA3TIP derived from these cell lines performs identically in vitro in CD2 receptor binding and T cell assays, examination of a pharmacodynamic marker-the reduction in CD2+ lymphocyte numbers-following the administration of equal doses of NS-0 or CHO derived LFA3TIP to baboons, suggested that the effect of the NS-0 derived material was less sustained. Pharmacokinetic analysis of the materials in baboons and mice shows that LFA3TIP produced by NS-0 cells is rapidly cleared from circulation relative to the product derived from CHO cells. The disparate clearance profiles correlate with distinct glycosylation patterns, with LFA3TIP derived from NS-0 cells being less extensively sialylated than that from CHO cells due in part to alpha-galactosyl capping of selected lactosamine moieties in the N-linked glycans of NS-0 derived LFA3TIP. Moreover, enzymatic desialylation of CHO derived LFA3TIP results in a glycoprotein with an evanescent serum profile when administered to mice and baboons. These results correlate the extent of N-acetylneuraminic acid capping with the clearance rates of LFA3TIP derived from the two distinct cell lines, and underscore the importance of evaluating glycosylation dependent PK parameters when choosing production cell lines for recombinant immunotherapeutics.
authors.affiliation: Biogen Inc., 14 Cambridge Center, MA 02142, USA.
authors.forename: W
authors.initials: W
authors.lastname: Meier
authors.forename: A
authors.initials: A
authors.lastname: Gill
authors.forename: M
authors.initials: M
authors.lastname: Rogge
authors.forename: R
authors.initials: R
authors.lastname: Dabora
authors.forename: G R
authors.initials: GR
authors.lastname: Majeau
authors.forename: F B
authors.initials: FB
authors.lastname: Oleson
authors.forename: W E
authors.initials: WE
authors.lastname: Jones
authors.forename: D
authors.initials: D
authors.lastname: Frazier
authors.forename: K
authors.initials: K
authors.lastname: Miatkowski
authors.forename: P S
authors.initials: PS
authors.lastname: Hochman
mesh_terms: D000276:Adjuvants, Immunologic / Q000096:biosynthesis / Q000493:pharmacokinetics* / Q000494:pharmacology*; D000818:Animals; D018801:CD2 Antigens / Q000235:genetics / Q000276:immunology; D018968:CD58 Antigens / Q000494:pharmacology*; D016466:CHO Cells / Q000378:metabolism; D006224:Cricetinae; D006031:Glycosylation; D006801:Humans; D007074:Immunoglobulin G / Q000494:pharmacology*; D008213:Lymphocyte Activation; D051379:Mice; D008822:Mice, Transgenic; D010215:Papio; D011134:Polysaccharides / Q000032:analysis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000493:pharmacokinetics* / Q000494:pharmacology*; D013601:T-Lymphocytes / Q000187:drug effects* / Q000276:immunology; D014407:Tumor Cells, Cultured / Q000378:metabolism
chemical_list: D000276:Adjuvants, Immunologic; D018801:CD2 Antigens; D018968:CD58 Antigens; D007074:Immunoglobulin G; D011134:Polysaccharides; D011993:Recombinant Fusion Proteins