A chromatographic approach for elevating the antibody-dependent cellular cytotoxicity of antibody composites.
Recent studies have shown that antibodies with low fucose content in their oligosaccharides exhibit highly potent antibody-dependent cellular cytotoxicity (ADCC). However, composites of therapeutic antibodies produced by conventional production systems using cell lines such as Chinese hamster ovary (CHO) and SP2/0 do not necessarily contain sufficient amounts of non-fucosylated antibody species. In this study, we combined two lectin-affinity chromatography techniques, Concanavalin A and Lens culinaris agglutinin, to enrich the non-fucosylated species from therapeutic material using the anti-Her2/neu model antibody. Oligosaccharide analysis by matrix-assisted laser desorption/ionization-time of flight MS following peptide-N-glycosidase F digestion suggested that non-fucosylated antibody could be enriched in the purified fraction with efficient removal of high-mannose species. The ADCC activity of the purified fraction was about 100-fold higher than that of the initial material. The chromatographic strategy presented here can be a useful tool to elevate ADCC activity of antibody materials without concentrating high-mannose oligosaccharides.
Tojo
Setsuko
S
Pharmaceutical Research Center, Kyowa Hakko Kirin Co., Ltd., Asahi-machi, Machida, Tokyo, Japan.
Okazaki
Akira
A
Wakitani
Masako
M
Shinkawa
Toyohide
T
Uchida
Kazuhisa
K
Suzawa
Toshiyuki
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D061067:Antibodies, Monoclonal, Humanized; D000920:Antibody-Dependent Cell Cytotoxicity; D001666:Binding Sites, Antibody; D016466:CHO Cells; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003208:Concanavalin A / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D003601:Cytotoxicity Tests, Immunologic / Q000379:methods*; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D008766:Methylmannosides / Q000276:immunology / Q000378:metabolism*; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000276:immunology / Q000378:metabolism*; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000276:immunology* / Q000378:metabolism; D000068878:Trastuzumab
D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D003208:Concanavalin A; D007074:Immunoglobulin G; D008766:Methylmannosides; D009844:Oligosaccharides; D018719:Erb-b2 Receptor Tyrosine Kinases; D000068878:Trastuzumab; C008466:methylmannoside; C508053:ERBB2 protein, human