title: Comparison of biological activity among nonfucosylated therapeutic IgG1 antibodies with three different N-linked Fc oligosaccharides: the high-mannose, hybrid, and complex types.
abstract: The structure of asparagine-linked oligosaccharides attached to the antibody constant region (Fc) of human immunoglobulin G1 (IgG1) has been shown to affect the pharmacokinetics and antibody effector functions of antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). However, it is still unclear how differences in the N-linked oligosaccharide structures impact the biological activities of antibodies, especially those lacking core fucose. Here, we succeeded in generating core fucose-lacking human IgG1 antibodies with three different N-linked Fc oligosaccharides, namely, a high-mannose, hybrid, and complex type, using the same producing clone, and compared their activities. Cultivation of an alpha-1,6-fucosyltransferase (FUT8) knockout Chinese hamster ovary cell line in the presence or absence of a glycosidase inhibitor (either swainsonine or kifunensine) yielded antibody production of each of the three types without contamination by the others. Two of three types of nonnaturally occurring atypical oligosaccharide IgG1, except the complex type, reduced the affinity for both human lymphocyte receptor IIIa (FcgammaRIIIa) and the C1q component of the complement, resulting in reduction of ADCC and CDC. The bulky structure of the nonreducing end of N-linked Fc oligosaccharides is considered to contribute the CDC change, whereas the structural change in the reducing end, i.e. the removal of core fucose, causes ADCC enhancement through improved FcgammaRIIIa binding. In the pharmacokinetic profile, although no significant difference of human neonatal Fc receptor (FcRn)-binding affinity was observed among the three types, the complex type showed longer serum half-lives than the other types irrespective of core fucosylation in mice, which also suggests the contribution of the nonreducing end structure. The present study provides basic information on the effects of core fucose-lacking N-linked Fc oligosaccharides on antibody biological activities.
authors.affiliation: Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd, 3-6-6 Asahi-machi, Machida-shi, Tokyo 194-8533, Japan.
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Kanda
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Yamada
authors.forename: Katsuhiro
authors.initials: K
authors.lastname: Mori
authors.forename: Akira
authors.initials: A
authors.lastname: Okazaki
authors.forename: Miho
authors.initials: M
authors.lastname: Inoue
authors.forename: Kazuko
authors.initials: K
authors.lastname: Kitajima-Miyama
authors.forename: Reiko
authors.initials: R
authors.lastname: Kuni-Kamochi
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Nakano
authors.forename: Keiichi
authors.initials: K
authors.lastname: Yano
authors.forename: Shingo
authors.initials: S
authors.lastname: Kakita
authors.forename: Kenya
authors.initials: K
authors.lastname: Shitara
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Satoh
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry / Q000493:pharmacokinetics / Q000627:therapeutic use; D058846:Antibodies, Monoclonal, Murine-Derived; D000920:Antibody-Dependent Cell Cytotoxicity / Q000502:physiology; D016466:CHO Cells; D002240:Carbohydrate Sequence; D015922:Complement C1q / Q000378:metabolism; D003165:Complement System Proteins / Q000378:metabolism / Q000502:physiology; D006224:Cricetinae; D003412:Cricetulus; D003602:Cytotoxicity, Immunologic / Q000502:physiology; D005260:Female; D005647:Fucosyltransferases / Q000235:genetics; D015395:Histocompatibility Antigens Class I / Q000378:metabolism; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000737:chemistry* / Q000627:therapeutic use; D007074:Immunoglobulin G / Q000737:chemistry* / Q000627:therapeutic use; D007155:Immunologic Factors / Q000737:chemistry* / Q000627:therapeutic use; D008351:Mannans / Q000737:chemistry*; D008657:Metabolic Clearance Rate; D051379:Mice; D008969:Molecular Sequence Data; D009101:Multiple Myeloma / Q000188:drug therapy; D009844:Oligosaccharides / Q000737:chemistry*; D030781:Organisms, Genetically Modified; D011485:Protein Binding; D011961:Receptors, Fc / Q000378:metabolism; D017452:Receptors, IgG / Q000378:metabolism; D000069283:Rituximab; D013329:Structure-Activity Relationship
chemical_list: D000911:Antibodies, Monoclonal; D058846:Antibodies, Monoclonal, Murine-Derived; C496124:FCGR3A protein, human; D015395:Histocompatibility Antigens Class I; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D007155:Immunologic Factors; D008351:Mannans; D009844:Oligosaccharides; D011961:Receptors, Fc; D017452:Receptors, IgG; C097323:polymannose; D000069283:Rituximab; D015922:Complement C1q; D003165:Complement System Proteins; D005647:Fucosyltransferases; C020559:Glycoprotein 6-alpha-L-fucosyltransferase; C100688:Fc receptor, neonatal