Epitope mapping and characterization of monoclonal antibodies to human protein C.
Six monoclonal antibodies specific to human protein C were characterized. Epitopes of these antibodies were determined on isolated proteolytic peptides of protein C by immunological methods. Three antibodies bound light chain of protein C: PC01 bound the gamma-carboxyglutamic acid domain calcium-dependently, while PC02 and PC08 bound the first epidermal growth factor-like domain in calcium-dependent and independent manners, respectively. The other three antibodies bound the heavy chain of protein C: PC13 bound activation peptide, PC04 recognized the activation site and PC09 bound the region close to a disulfide bond connecting light and heavy chains. Activation of protein C with thrombin-thrombomodulin complex was inhibited strongly by PC04 and moderately by PC08, PC09 and PC13. PC04 and PC13 may directly block the activation site. On the other hand, epitopes of PC08 and PC09 may be involved in interaction between protein C and thrombin-thrombomodulin complex, or locate close to activation site on the tertiary structure of protein C. Anticlotting activity of protein C was inhibited strongly by PC01 and moderately by PC02, PC08 and PC09, while amidolytic activity was inhibited only by PC09. The epitopes described here may constitute part of protein-C-specific sites, which are important for the function of protein C.
Takahashi
M
M
Pharma Research Laboratories, Hoechst Japan Ltd., Saitama.
Kimura
M
M
Kurosawa-Ohsawa
K
K
Iwaki
A
A
Tanaka
S
S
Matsuishi
T
T
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D001667:Binding, Competitive; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D004789:Enzyme Activation; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D011486:Protein C / Q000276:immunology* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000939:Epitopes; D011486:Protein C