Flow and antibody binding properties of hydrated fibrins prepared from plasma, platelet rich plasma and whole blood.
Previous studies, using cross-linked fibrin prepared from purified fibrinogen, showed low binding of a fibrin-specific monoclonal antibody designated T2G1 (Procyk et al., Blood 77:1469-75, 1991). In this study we investigated the binding of T2G1 and one other antibody to clots prepared from platelet poor plasma (PPP), platelet rich plasma (PRP) and whole blood. In contrast to our previous study, we used unlabelled antibodies and quantitated the level bound by ELISA, measuring antibody concentration in the non-adsorbed fraction. Antibody T2G1 bound 1.35 +/- 0.10 pmol/pmol fibrin (n = 11) to whole blood columns, 1.64 +/- 0.18 (n = 10) to PRP columns and 1.58 +/- 0.13 (n = 8) to PPP columns. The binding of T2G1 to columns made from purified fibrinogen was 0.78 +/- 0.05 pmol/pmol fibrin (n = 15). An antibody to a conformation-dependent epitope on Fragment D (Fd4-7B3) bound in comparable amounts to the different fibrins. Flow data show that whole blood columns, and also, but to a lesser extent those made with plasma, had a higher flow rate, permeability and fiber mass-length ratio than columns prepared from fibrinogen indicating a more coarse fibrin network. These data show that the presence of other proteins and blood cells, similar to what might occur in vivo, not only lead to an increase in the permeability of gels but also allow for better exposure of some epitopes.
Bini
A
A
Lindsley F. Kimball Research Institute, New York Blood Center, NY 10021.
Callender
S
S
Procyk
R
R
Blombäck
B
B
Kudryk
B J
BJ
D000911:Antibodies, Monoclonal / Q000276:immunology*; D001769:Blood; D000939:Epitopes / Q000276:immunology; D005337:Fibrin / Q000276:immunology / Q000302:isolation & purification*; D005782:Gels; D006801:Humans; D010949:Plasma; D011487:Protein Conformation
D000911:Antibodies, Monoclonal; D000939:Epitopes; D005782:Gels; D005337:Fibrin