Neutralization of the antiheparin activity of platelet factor 4 by a monoclonal antibody.
We have produced a panel of monoclonal antibodies (mAbs) against rabbit platelet factor 4 (PF4). Two of these mAbs have been characterized in this study. In particular the antibody called 10B2, which also recognizes the human molecule, is able to block PF4's ability to neutralize heparin in a modified Heparin-Factor Xa chromogenic assay. The inhibition appears to be more than 95% at 1:1 mAb/PF4 molar ratio both for purified rabbit and human PF4. Similar results were obtained using supernatants from stimulated human platelets (90% of inhibition at 1:1 mAb/PF4 molar ratio) or using Fab fragments from 10B2. Studies to determine the antigenic determinant against which 10B2 is directed, show that this is an assembled epitope which involves disulfide bonds of the PF4.
Saggin
L
L
Dept. of Vascular Biology, Division, FIDIA S.p.A., Abano Terme, Italy.
Cazzola
F
F
Corona
G
G
Salvatico
E
E
Cella
G
G
Prosdocimi
M
M
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001665:Binding Sites; D001792:Blood Platelets / Q000276:immunology; D000939:Epitopes / Q000737:chemistry; D006494:Heparin Antagonists / Q000276:immunology*; D006801:Humans; D007140:Immunoglobulin Fab Fragments; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D009842:Oligopeptides / Q000737:chemistry / Q000276:immunology; D010978:Platelet Factor 4 / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000276:immunology*; D011817:Rabbits
D000911:Antibodies, Monoclonal; D000939:Epitopes; D006494:Heparin Antagonists; D007140:Immunoglobulin Fab Fragments; D009842:Oligopeptides; D010978:Platelet Factor 4