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Sci Rep. 2020 Jan 30;10(1):1464. doi: 10.1038/s41598-019-57393-9.

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

Author information

1
Bristol Institute for Transfusion Sciences, International Blood Group Reference Laboratory, NHS Blood and Transplant, Bristol, UK. bm.kumpel@btinternet.com.
2
NIBRT Glycoscience Group, National Institute for Bioprocessing Research and Training, Dublin, Ireland.
3
Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands.
4
Department of Pathology, University of Cambridge, Cambridge, UK.
5
National Research Center for Hematology, Moscow, Russia.
6
Sanquin Research and Landsteiner Laboratory, Department for Experimental Immunohematology, Academic Medical Centre, University of Amsterdam, Amsterdam, The Netherlands. G.Vidarsson@sanquin.nl.
7
Sanquin Research and Landsteiner Laboratory, Department for Experimental Immunohematology, Academic Medical Centre, University of Amsterdam, Amsterdam, The Netherlands.

Abstract

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 <50% fucosylation mediated more efficient ADCC and clearance than anti-D Ig. Galactosylation of B mAb-Ds was 57-83% but 15-58% for rodent mAb-Ds. HH mAb-Ds had non-human sugars. These data reveal high galactosylation like anti-D Ig (>60%) together with lower fucosylation (<60%) as safe features of mAb-Ds for mediating rapid red cell clearance at low doses, to enable effective, inexpensive prophylaxis.

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