Isolation and characterization of monoclonal antibodies to a recombinant human topoisomerase II polypeptide.
We have produced two murine monoclonal antibodies (SWT3D1 and SWR1C2) to a recombinant polypeptide corresponding to the carboxyl-terminal one-third (amino acid 854-amino acid 1447) of human topoisomerase II alpha. Each antibody is able to recognize intact human topoisomerase II using immunoblotting and enzyme-linked immunosorbent assay (ELISA) techniques. Data is presented demonstrating that the antibodies bind specifically to topoisomerase II alpha but do not interact with topoisomerase II beta. The monoclonal antibodies do not recognize murine or calf thymus topoisomerase II indicating that each may bind exclusively to the human enzyme. The topoisomerase II binding sites for each monoclonal antibody have been compared in a competition ELISA. The SWT3D1 antibody had no significant effect on the binding efficiency of biotinylated SWR1C2 antibody. Although SWR1C2 was capable of inhibiting the binding of biotinylated SWT3D1, this only occurred at concentrations approximately 1000-fold higher than those required of SWT3D1 to block binding of itself. These results suggest that SWT3D1 and SWR1C2 do not recognize identical epitopes on topoisomerase II.
Robinson
R G
RG
Sterling Winthrop Pharmaceuticals Research Division, Sterling Winthrop Inc., Collegeville, PA 19426.
Rapp
L
L
Bowdish
K J
KJ
Graham
M A
MA
Huff
A C
AC
Coughlin
S A
SA
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification*; D015153:Blotting, Western; D004250:DNA Topoisomerases, Type II / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007527:Isoenzymes / Q000276:immunology; D051379:Mice; D009363:Neoplasm Proteins / Q000276:immunology; D010446:Peptide Fragments / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D007527:Isoenzymes; D009363:Neoplasm Proteins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D004250:DNA Topoisomerases, Type II