Accessibility of epitopes on fibrin clots and fibrinogen gels.
Radiolabeled antibodies were perfused into fibrin clots and fibrinogen gels formed in vitro to assess the reactivity of selected epitopes. An antifibrinogen monoclonal antibody (MoAb) (antibody 1D4/xl-f), directed against an epitope in the A alpha-chain C-terminal region (A alpha 241-476), bound to 35% of the epitope in crosslinked fibrin clots and 37% of the same epitope in factor XIII-induced fibrinogen gel networks. A different MoAb (4-2/xl-f, anti gamma 392-406) bound to only 7% of the epitope in both fibrin and fibrinogen gels. As expected, an antifibrin MoAb (antibody T2G1, antiB beta 15-21) did not bind to fibrinogen gels, but bound to fibrin, although to only 14% of the available T2G1-reactive epitopes. An antibody that does not recognize fibrin (antibody 1-8C6, antiB beta 1-21) predictably did not bind to fibrin clots and bound to 35% of the 1-8C6 epitopes present in fibrinogen gels, a level of binding also observed with antibody T2G1 and fibrinogen gels only after the latter were treated with thrombin. T2G1 epitope expression was affected much more than 1D4/xl-f epitope expression in clots formed in buffers of high or low ionic strength, conditions known to influence clot structure. Studies on the availability, in quantitative terms, of the T2G1-reactive epitope in fibrin clots is of particular importance because this antibody is currently being used in clinical trials as a clot imaging agent.
Procyk
R
R
Department of Blood Coagulation Biochemistry, New York Blood Center, NY 10021.
Kudryk
B
B
Callender
S
S
Blombäck
B
B
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D001777:Blood Coagulation; D000939:Epitopes / Q000032:analysis*; D005337:Fibrin / Q000276:immunology / Q000502:physiology*; D005340:Fibrinogen / Q000276:immunology / Q000502:physiology*; D005782:Gels; D006801:Humans; D007700:Kinetics; D008024:Ligands
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000939:Epitopes; D005782:Gels; D008024:Ligands; D005337:Fibrin; D005340:Fibrinogen