title: Lipopolysaccharide epitope specificity and binding cross reactivity of the human IgM anti-lipid A monoclonal antibody SdJ5-1.17.15.
abstract: The binding properties and specificity of the SdJ5-1.17.15 (SdJ5) human IgM monoclonal antibody (mAb), prepared against S. minnesota R595 heat killed whole organisms, were assessed by dot blot assay in vitro. In that assay, lipopolysaccharide (LPS) related antigens, immobilized on nitrocellulose membrane, were reacted with the test and control human IgM antibodies. Results indicated that SdJ5 mAb reacted specifically with lipid A, a component of LPS, from a variety of bacterial species, but not with the whole LPS molecule. The inability of the mAb to react with whole LPS in dot blot assay was attributed to the possible effect of the solid phase on epitope exposure or structure. This hypothesis was tested by inhibition studies which indicated that liquid phase adsorption with LPS abolished or greatly reduced the specific recognition of solid phase lipid A by the mAb. These results indicated that LPS-associated antigens were recognized by the SdJ5 anti-lipid A mAb in liquid, but not solid, phase. In an attempt to identify the SdJ5-specific epitope on lipid A, the pattern of reactivities of different lipid A analogues with the mAb were examined by dot blot assay. Results indicated that a combination of the fatty acid side chains and the phosphate groups of lipid A (both groups being implicated as important for sepsis mediation) were either directly involved in the epitope structure or affected the exposure of epitope in solid phase. Because SdJ5 recognizes an LPS epitope on lipid A closely associated with endotoxin activity, this mAb could potentially be a useful therapeutic agent against septic shock.
authors.affiliation: Baxter Healthcare Corporation, Biotech Group, Hyland Division, Duarte, CA 91010.
authors.forename: M
authors.initials: M
authors.lastname: Kazemi
authors.forename: C C
authors.initials: CC
authors.lastname: Huntenburg
authors.forename: J E
authors.initials: JE
authors.lastname: Bubbers
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions / Q000276:immunology; D003429:Cross Reactions; D004306:Dose-Response Relationship, Immunologic; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D006090:Gram-Negative Bacteria / Q000276:immunology; D006801:Humans; D007075:Immunoglobulin M / Q000276:immunology*; D008050:Lipid A / Q000737:chemistry / Q000276:immunology*; D008070:Lipopolysaccharides / Q000276:immunology*; D015394:Molecular Structure; D010449:Peptide Mapping
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D007075:Immunoglobulin M; D008050:Lipid A; D008070:Lipopolysaccharides