Production and characterization of monoclonal antibodies against bovine pancreatic trypsin inhibitor.
Two monoclonal antibodies (MAbs) have been produced, without the use of a supporting carrier, against bovine basic pancreatic trypsin inhibitor (BPTI or aprotinin), a mini-protein composed of 58 amino acids. Both MAbs obtained were found to be IgM. One of them was purified and further characterized. This MAb (ICI) binds to the immunogen with an association constant of 1.6 X 10(6)M-1 at pH 7.4. Competition experiments with trypsin or inactivated trypsin demonstrate that ICI MAb interacts with BPTI at, or near, the proteinase-binding site. ICI MAb binds, with a much lower association constant (approximately 200M-1), to an isoinhibitor (spleen inhibitor II) which differs from BPTI in seven amino-acids; three of these substitutions are at the active site, in the contact area with the proteinase.
Marini
S
S
Dipartimento di Medicina Sperimentale e Scienze Biochimiche, Università Tor Vergata, Roma.
Fiorucci
L
L
Fioretti
E
E
Giardina
B
B
Ascoli
F
F
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000096:biosynthesis* / Q000276:immunology; D000915:Antibody Affinity; D000936:Antigen-Antibody Complex / Q000378:metabolism*; D007611:Aprotinin / Q000031:analogs & derivatives*; D002417:Cattle; D007075:Immunoglobulin M / Q000032:analysis; D014359:Trypsin Inhibitor, Kazal Pancreatic / Q000276:immunology*; D014361:Trypsin Inhibitors / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D007075:Immunoglobulin M; D014361:Trypsin Inhibitors; C062659:spleen protease inhibitor II; D014359:Trypsin Inhibitor, Kazal Pancreatic; D007611:Aprotinin