Analysis of idiotope structure of ovarian cancer antibodies: recognition of the same epitope by two monoclonal antibodies differing mainly in their heavy chain variable sequences.
Two MoAbs, independently raised against ovarian carcinoma cells and referred to as OV-TL3 and OV-TL16, display an identical reaction pattern with a membrane-associated protein in both normal and malignant ovarian cells. Also, a similar binding affinity constant and a similar number of binding sites per cell indicate that both MoAbs bind to the same antigen. Competition assays reveal that OV-TL16 is able to compete with OV-TL3 for binding to OVCAR-3 cells. Epitope mapping using a filamentous phage hexapeptide epitope library showed that both MoAbs are able to select identical phages, suggesting that their epitopes are identical or at least overlapping. However, purified polyclonal and monoclonal anti-idiotypic antibodies directed against OV-TL3 failed to recognize the OV-TL16 idiotype, indicating that the structure of the antigen-binding regions of both antibodies is distinct. This was corroborated by molecular cloning and sequencing of the variable heavy (VH) and light (VL) chain immunoglobulin regions of both MoAbs. The VH regions of both antibodies were found to be distinct, whereas the VL regions are almost identical. Computer modelling of the idiotypes suggests that the complementarity determining regions (CDR), with the exception of VHCDR3, have (almost) identical spatial configurations. Our data indicate that, although structurally different in their VH regions, OV-TL3 and OV-TL16 are able to bind to identical epitopic regions on the antigen, because differences in primary structure do not exclude the formation of sufficient and similar spatial structures for the interaction with an epitope.
Slobbe
R
R
Department of Molecular Cell Biology and Genetics, University of Limburg, Maastricht, The Netherlands.
Poels
L
L
ten Dam
G
G
Boerman
O
O
Nieland
L
L
Leunissen
J
J
Ramaekers
F
F
van Eys
G
G
D000595:Amino Acid Sequence; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology; D000912:Antibodies, Neoplasm / Q000276:immunology*; D001483:Base Sequence; D003198:Computer Simulation; D003429:Cross Reactions / Q000276:immunology; D018604:Epitope Mapping; D000939:Epitopes / Q000276:immunology; D005260:Female; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000276:immunology; D018448:Models, Immunological; D008969:Molecular Sequence Data; D010051:Ovarian Neoplasms / Q000276:immunology*; D010053:Ovary / Q000276:immunology; D014407:Tumor Cells, Cultured
D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D000912:Antibodies, Neoplasm; D000939:Epitopes; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains