Reactivity with legume lectins of three monoclonal antibodies made against the Lathyrus odoratus lectin.
Three murine IgG1 monoclonal antibodies (MoAb) were produced against the glucose/mannose-specific two-chain mitogen from seeds of Lathyrus odoratus (sweet pea) belonging to the Vicieae tribe of the Leguminosae family. Their antigenic specificities were tested against subunits of two-chain and one-chain legume lectins separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and electrotransferred to nitrocellulose filters. Different binding to native and detergent-treated lectins in dot-blots indicated that two of the antibodies bound to conformational epitopes which by immunoblotting were shown to be expressed on the heavy subunits from all the five tested two-chain lectins and on the six one-chain lectins, including PHA (Phaseolus vulgaris) and Con A (Canavalia ensiformis). However, ELISA cross-inhibition studies showed that these two antibodies bound to different epitopes on the lectin molecules. The third MoAb reacted with a continuous epitope not revealed by classical taxonomy since only three of five heavy subunits of the two-chain lectins stained with the antibody. Comparison of the known amino-acid sequences of the chains indicates eight positions as possible binding sites for this antibody.
Kolberg
J
J
Department of Immunology, National Institute of Public Health, Oslo.
Michaelsen
T E
TE
Wedege
E
E
Heier
H E
HE
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex / Q000032:analysis; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D007887:Fabaceae; D037102:Lectins / Q000276:immunology*; D008970:Molecular Weight; D010835:Phytohemagglutinins / Q000276:immunology*; D037121:Plant Lectins; D010946:Plants, Medicinal; D013045:Species Specificity
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D037102:Lectins; D010835:Phytohemagglutinins; D037121:Plant Lectins; C039412:lathyrus lectin