Monoclonal antibodies against rice bran lectin.
Two mouse monoclonal antibodies (IgM) were produced against the N-acetylglucosamine-specific rice bran lectin. It was difficult to establish antibody-producing hybrids when soluble rice lectin was used for immunization. Therefore a complex of rice lectin and chitin, a water-insoluble polymer of N-acetylglucosamine, was used as immunogen. Antibody reactivity against Gramineae lectins from barley, rice, rye and wheat (wheat germ agglutinin) was tested in ELISA and two (137,E-1 and 140,B-3) were found to be specific for rice lectin. Gramineae lectins were separated by sodium dodecyl-sulphate-polyacrylamide gel electrophoresis and electroblotted to nitrocellulose papers. Analyses showed that the antibodies reacted strongly with non-reduced rice lectin, whereas only weak staining was seen with the other lectins. The binding was abolished after treatment with 2-mercaptoethanol suggesting that disulphide bond tertiary structures were necessary for epitope integrity.
Kolberg
J
J
Department of Immunology, National Institute of Public Health, Oslo, Norway.
D000117:Acetylglucosamine / Q000032:analysis; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D002523:Edible Grain; D004797:Enzyme-Linked Immunosorbent Assay; D037102:Lectins / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D008969:Molecular Sequence Data; D012275:Oryza; D037121:Plant Lectins; D012689:Sequence Homology, Nucleic Acid
D000911:Antibodies, Monoclonal; D037102:Lectins; D037121:Plant Lectins; C040599:rice lectin; D000117:Acetylglucosamine