Production and characterization of two monoclonal antibodies to human pancreatic trypsin 1.
Two monoclonal antibodies (MAb) G6 and A8 directed against human trypsin 1 have been produced by hybridization of myeloma cells with spleen cells of OF1 immunized mice. Antibodies were screened by radioimmunoassay. Monoclonal antibodies were purified by affinity chromatography on Protein A-Sepharose, and we found that MAb G6 was of the IgG2b class and MAb A8 of the IgG2a class. Both MAbs had a high affinity for trypsin 1 with the respective affinity constants equal to 1.3 x 10(8) l/mol for G6 and 3.2 x 10(7) l/mol for A8. Epitope specificity was studied by western blotting, using human trypsinogens 1 and 2. Both MAbs gave a positive reaction with native trypsinogen 1 and no reaction with the same protein after reduction. Only MAb G6 reacted with trypsinogen 2 in the native form. Its affinity for trypsin 2 was found similar to that for trypsin 1 with a constant equal to 2.7 x 10(7) l/mol. Both antibodies appeared directed against conformational and not sequential epitopes.
Lafont
P
P
Faculté de Médecine Lyon-Sud, Oullins, France.
Paulin
C
C
Brayle
A
A
Figarella
C
C
Barbier
Y
Y
Guy-Crotte
O
O
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000302:isolation & purification; D000915:Antibody Affinity; D000918:Antibody Specificity; D015153:Blotting, Western; D006801:Humans; D006825:Hybridomas; D051379:Mice; D010189:Pancreatic Juice / Q000201:enzymology*; D014357:Trypsin / Q000276:immunology*; D014362:Trypsinogen / Q000276:immunology
D000911:Antibodies, Monoclonal; D014362:Trypsinogen; D014357:Trypsin