Production and characterization of monoclonal antibodies to human pancreatic elastase 2.
Three murine monoclonal antibodies (MAbs) against human pancreatic elastase 2 (ELS-2) were produced by hybridomas obtained from the fusion of murine myeloma cells, P3X63-Ag8.-653, with the splenocytes of mice immunized with recombinant human proELS-2 (r-proELS-2) and purified natural ELS-2. These MAbs were found to recognize the different epitopes of ELS-2 based on a competitive binding assay. In addition, one of the MAbs, E19, reacted with the activation peptide of ELS-2. Western blot analysis performed under nonreducing conditions indicated that all MAbs bound only to ELS-2 in pancreatic juice in addition to natural ELS-2. Reduced ELS-2 on a Western blot could not be detected with E19, indicating that the activation peptide remained attached to active ELS-2 via a disulfide bond even after tryptic activation. Another MAb, B4, inhibited the enzymatic activity of ELS-2, indicating that B4 recognized the active site or its vicinity to ELS-2.
Shirasu
Y
Y
Pharmaceutical Laboratory, Kirin Brewery Co., Ltd., Gunma, Japan.
Shimada
Y
Y
Takemura
K
K
Izumi
T
T
Miyazaki
H
H
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D001665:Binding Sites; D001667:Binding, Competitive; D000939:Epitopes / Q000276:immunology; D006801:Humans; D007120:Immunochemistry; D051379:Mice; D009500:Neutralization Tests; D010179:Pancreas / Q000201:enzymology*; D010196:Pancreatic Elastase / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D010196:Pancreatic Elastase