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. 2019 Aug 27;10:1967. doi: 10.3389/fimmu.2019.01967

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Copyright © 2019 Shathili, Bandala-Sanchez, John, Goddard-Borger, Thaysen-Andersen, Everest-Dass, Adams, Harrison and Packer.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Comparative N-glycoprofiling of recombinant human IgG Fc and CD52. (A) Proliferation of human PBMCs (³H thymidine uptake) followed 5 days incubation with tetanus toxoid (10 LfU), histograms show mean ± SD of within-assay triplicates, in the presence of different concentration of proteins (CD52-Fc 5, 10, 50 μg/ml; Cleaved CD52-Fc 50 μg/ml and Fc control 50 μg/ml). The Fc component was cleaved from CD52-Fc with Factor Xa. (B) Factor Xa treated-CD52 was analyzed by Western blotting with anti-CD52-HRP antibody (Campath-H1). (C) Summed MS profile of N-glycans released from the Fc (I) and CD52 (II); the latter variant was generated by introducing a point mutation (A297N) into the conventional Fc N-glycosylation site.