Open AccessMonoclonal Antibodies Against Fusicoccin with Binding Characteristics Similar to the Putative Fusicoccin Receptor of Higher Plants
Author(s) -
Martin Feyerabend,
Elmar W. Weiler
Publication year - 1987
Publication title -
plant physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.554
H-Index - 312
eISSN - 1532-2548
pISSN - 0032-0889
DOI - 10.1104/pp.85.3.835
Subject(s) - fusicoccin , immunogen , monoclonal antibody , chemistry , cardenolide , microbiology and biotechnology , hapten , antiserum , antibody , immunoassay , biochemistry , biology , chromatography , stereochemistry , enzyme , glycoside , immunology , atpase
Monoclonal antibodies were raised against fusicoccin. The toxin, linked to bovine serum albumin through its t-pentenyl moiety, served as immunogen. Hybridomas secreting anti-fusicoccin antibodies were screened by radioimmunoassay employing a novel radioactive derivative, [(3)H]-nor-fusicoccin-alcohol of high specific activity (1.5 x 10(14)Bq/mole). The two monoclonal antibodies reported here are of high apparent affinity for fusicoccin (0.71 x 10(-9) molar and 1.85 x 10(-9) molar). This is comparable to the apparent affinity of rabbit antiserum raised against the same type of conjugate (9.3 x 10(-9) molar). A method for the single step purification of the monoclonal antibodies from ascites fluid is reported. A solid-phase immunoassay, using alkaline phosphatase as enzyme, exhibits a measuring range from 0.1 to 1.5 picomoles (about 70 picograms to 1 nanogram) of fusicoccin. The displacement of [(3)H]-nor-fusicoccin-alcohol from the antibodies by compounds structurally related to fusicoccin exhibits similar selectivity as a microsomal binding assay with the same tracer as radiolabeled probe.