Postadsorptive transitions in fibrinogen adsorbed to biomer: changes in baboon platelet adhesion, antibody binding, and sodium dodecyl sulfate elutability.
Residence-time-dependent changes in fibrinogen after its adsorption to Biomer were examined by measuring platelet adhesion and antibody binding to the adsorbed protein, and the amount of adsorbed fibrinogen which could be eluted by sodium dodecyl sulfate (SDS). Baboon fibrinogen was first adsorbed (from either pure solution or dilute plasma) to Biomer, which was then stored in either buffer or buffered albumin solution prior to testing. Subsequently, the adherent protein layer was either probed for fibrinogen capable of mediating platelet adhesion using 111In radiolabeled, washed platelet suspensions under both static and shearing conditions, or for fibrinogen capable of binding antibody using a direct enzyme linked immunosorbent assay (ELISA). Alternatively, the surface with the adsorbed protein layer was soaked in a 3% SDS solution, and the amount of 125I radiolabeled fibrinogen retained was measured. Decreases in platelet and antibody binding, and in the SDS elutability of the adsorbed fibrinogen after it was stored in buffer were detected, although different rates of decrease were observed for each method. When the protein-coated surfaces were stored in buffered albumin solution rather than buffer, the decrease in the reactivity of fibrinogen was prevented. While each of the three assays measures a different property of adsorbed fibrinogen, this study suggests that the adherent protein undergoes time dependent conformational changes which render it less reactive toward platelets and antibodies, and more resistant to elution by SDS.
Chinn
J A
JA
Department of Chemical Engineering, University of Washington, Seattle 98195.
Posso
S E
SE
Horbett
T A
TA
Ratner
B D
BD
D000327:Adsorption; D000818:Animals; D000937:Antigen-Antibody Reactions / Q000187:drug effects*; D002021:Buffers; D004797:Enzyme-Linked Immunosorbent Assay; D005340:Fibrinogen / Q000737:chemistry* / Q000276:immunology; D066298:In Vitro Techniques; D008968:Molecular Conformation; D010215:Papio; D010973:Platelet Adhesiveness / Q000187:drug effects*; D011140:Polyurethanes / Q000737:chemistry*; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry
D002021:Buffers; D011140:Polyurethanes; D012967:Sodium Dodecyl Sulfate; C031892:biomer; D005340:Fibrinogen