A close look at human IgG sialylation and subclass distribution after lectin fractionation.
Polyspecific human IgG preparations are indicated for the treatment of primary immunodeficiency disorders associated with defects in humoral immunity. In addition, intraveneous IgG (IVIG) is used to treat patients with autoimmune and systemic inflammatory diseases. Lectin chromatography on Sambucus nigra agglutinin stood at the cradle of the hypothesis that the anti-inflammatory properties depend on sialylation of the N-glycans in the Fc region of IgG. A detailed analysis of fractions obtained by lectin chromatography revealed that binding of IVIG is essentially mediated by Fab glycosylation. Moreover, experiments with a monoclonal antibody from a human cell line and IVIG Fc fragments indicated that at least two sialic acids in the Fc region of an antibody are required for lectin binding. Such glycoforms contain either two monosialylated glycans or a disialylated glycan and constitute 1% or less of the total human IgG. Arguably this small proportion holds the entire anti-inflammatory potency. A new mass spectrometric quantification method of IgG subclass ratio revealed that the IVIG Fc preparation essentially consists of IgG1. This observation may be relevant when studying the effect of human Fc in murine models of inflammation because mouse IgG subclasses differ substantially in their interaction with receptors.
Stadlmann
Johannes
J
Department of Chemistry, University of Natural Resources and Applied Life Sciences (BOKU), Vienna, Austria.
Weber
Alfred
A
Pabst
Martin
M
Anderle
Heinz
H
Kunert
Renate
R
Ehrlich
Hartmut J
HJ
Peter Schwarz
Hans
H
Altmann
Friedrich
F
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western; D005591:Chemical Fractionation; D004591:Electrophoresis, Polyacrylamide Gel; D006031:Glycosylation; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000276:immunology; D007141:Immunoglobulin Fc Fragments / Q000737:chemistry / Q000276:immunology; D016756:Immunoglobulins, Intravenous / Q000737:chemistry* / Q000145:classification* / Q000276:immunology; D013058:Mass Spectrometry; D018448:Models, Immunological; D008969:Molecular Sequence Data; D037121:Plant Lectins / Q000276:immunology*; D054788:Ribosome Inactivating Proteins / Q000276:immunology*; D012685:Sepharose; D012794:Sialic Acids / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D007141:Immunoglobulin Fc Fragments; D016756:Immunoglobulins, Intravenous; D037121:Plant Lectins; C050600:Sambucus nigra lectins; D012794:Sialic Acids; D012685:Sepharose; D054788:Ribosome Inactivating Proteins