analysis and functional consequences of increased fab-sialylation of intravenous immunoglobulin (ivig) after lectin fractionation分析和功能的后果增加fab-sialylation凝集素分离后的静脉注射免疫球蛋白(丙种球蛋白).pdfVIP

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2017-09-07 发布于上海
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analysis and functional consequences of increased fab-sialylation of intravenous immunoglobulin (ivig) after lectin fractionation分析和功能的后果增加fab-sialylation凝集素分离后的静脉注射免疫球蛋白(丙种球蛋白).pdf

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analysis and functional consequences of increased fab-sialylation of intravenous immunoglobulin (ivig) after lectin fractionation分析和功能的后果增加fab-sialylation凝集素分离后的静脉注射免疫球蛋白(丙种球蛋白)

Analysis and Functional Consequences of Increased Fab- Sialylation of Intravenous Immunoglobulin (IVIG) after Lectin Fractionation ¨ 1 2 ¨ 1 1 1 Fabian Kasermann *, David J. Boerema , Monika Ruegsegger , Andreas Hofmann , Sandra Wymann , Adrian W. Zuercher1, Sylvia Miescher 1 1 Research Development, CSL Behring AG, Bern, Switzerland, 2 Research Development, CSL Behring LLC, Kankakee, Illinois, United States of America Abstract It has been proposed that the anti-inflammatory effects of intravenous immunoglobulin (IVIG) might be due to the small fraction of Fc-sialylated IgG. In this study we biochemically and functionally characterized sialic acid-enriched IgG obtained by Sambucus nigra agglutinin (SNA) lectin fractionation. Two main IgG fractions isolated by elution with lactose (E1) or acidified lactose (E2) were analyzed for total IgG, F(ab’)2 and Fc-specific sialic acid content, their pattern of specific antibodies and anti-inflammatory potential in a human in vitro inflammation system based on LPS- or PHA-stimulated whole blood. HPLC and LC-MS testing revealed an increase of sialylated IgG in E1 and more substantially in the E2 fraction. Significantly, the increased amount of sialic acid residues was primarily found in the Fab region whereas only a minor increase was observed in the Fc region. This indicates preferential binding of the Fab sialic acid to SNA. ELISA analyses of a representative range of pathogen and auto-antigens indicated a skewed antibody pattern of the sialylated IVIG fractions. Finally, the E2 fraction exerted a more profound anti-inflammatory effect compared to E1 or IVIG, evidenced by reduced CD54 expression on monocytes