Anti-D monoclonal antibodies from 23 human and rodent cell lines display diverse IgG Fc-glycosylation profiles that determine their clinical efficacy

• Published in: Scientific reports 2020-01, Vol.10 (1), p.1464-1464, Article 1464
• Main Authors: Kumpel, Belinda M., Saldova, Radka, Koeleman, Carolien A. M., Abrahams, Jodie L., Ederveen, Agnes Hipgrave, Armour, Kathryn L., Olovnikova, Natalia I., Vidarsson, Gestur, Kapur, Rick, Rudd, Pauline M., Wuhrer, Manfred
• Format: Article
• Language: English
• Subjects: 101/47; 13/1; 13/106; 13/62; 631/250/251/1574; 631/45/221; 631/61/475; 692/308/153; 692/699/1541; 82/58; Acids; Animals; Antibodies, Monoclonal - immunology; Antibodies, Monoclonal - metabolism; Antibodies, Monoclonal - therapeutic use; Antigens; Blood & organ donations; Cell Line; Clinical trials; Cloning; Cricetulus; Disease prevention; Erythroblastosis, Fetal - therapy; Fetuses; Fucose - metabolism; Galactose - metabolism; Glycosylation; High-performance liquid chromatography; Humanities and Social Sciences; Humans; Hybridomas - immunology; Immunization; Immunoglobulin G; Immunoglobulin G - immunology; Immunoglobulin G - metabolism; Laboratories; Liquid chromatography; Lymphoblastoid cell lines; Mass spectrometry; Mass spectroscopy; Mice; Monoclonal antibodies; multidisciplinary; N-Acetylneuraminic Acid - metabolism; Oligosaccharides; Ovaries; Prophylaxis; Rats; Rho(D) Immune Globulin - immunology; Rho(D) Immune Globulin - metabolism; Rho(D) Immune Globulin - therapeutic use; Rodents; Science; Science (multidisciplinary); Scientific imaging; Treatment Outcome
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-019-57393-9

Anti-D immunoglobulin (Anti-D Ig) prophylaxis prevents haemolytic disease of the fetus and newborn. Monoclonal IgG anti-Ds (mAb-Ds) would enable unlimited supplies but have differed in efficacy in FcγRIIIa-mediated ADCC assays and clinical trials. Structural variations of the oligosaccharide chains of mAb-Ds are hypothesised to be responsible. Quantitative data on 12 Fc-glycosylation features of 23 mAb-Ds (12 clones, 5 produced from multiple cell lines) and one blood donor-derived anti-D Ig were obtained by HPLC and mass spectrometry using 3 methods. Glycosylation of mAb-Ds from human B-lymphoblastoid cell lines (B) was similar to anti-D Ig although fucosylation varied, affecting ADCC activity. In vivo , two B mAb-Ds with 77–81% fucosylation cleared red cells and prevented D-immunisation but less effectively than anti-D Ig. High fucosylation (>89%) of mouse-human heterohybridoma (HH) and Chinese hamster ovary (CHO) mAb-Ds blocked ADCC and clearance. Rat YB2/0 mAb-Ds with 60%) together with lower fucosylation (