Production and characterisation of monoclonal antibodies against native and disassembled human catalase.
Catalase isolated from human erythrocytes was used to immunise mice, in order to generate hybridomas producing specific monoclonal antibodies to the enzyme. Hybridomas secreting anti-(catalase) antibodies were identified by a modified enzyme-linked immunosorbent assay (ELISA) using either monomer/dimer catalase or native, tetrameric enzyme. Three stable hybridoma clones were selected and the characteristics of the antibodies produced were investigated by ELISA, immunofluorescence, immunoprecipitation and immunoblotting experiments. One monoclonal antibody (17E10) was shown to interact with both native tetramer catalase and--to a lesser extent--with monomer/dimer catalase. Two monoclonal antibodies (10B12H9, 13A10) were found to react only with completely denatured catalase or with monomer/dimer catalase but not with native catalase.
Wiemer
E A
EA
E.C. Slater Institute for Biochemical Research, University of Amsterdam, Netherlands.
Ofman
R
R
Middelkoop
E
E
de Boer
M
M
Wanders
R J
RJ
Tager
J M
JM
D000911:Antibodies, Monoclonal / Q000276:immunology*; D002374:Catalase / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D046911:Macromolecular Substances; D015394:Molecular Structure; D011487:Protein Conformation; D013329:Structure-Activity Relationship
D000911:Antibodies, Monoclonal; D046911:Macromolecular Substances; D002374:Catalase