title: Residence time effects on monoclonal antibody binding to adsorbed fibrinogen.
abstract: Fibrinogen adsorbed to polymeric surfaces and then allowed to reside on the surface while it is kept in a buffer solution for a period of time (the 'residence time') undergoes postadsorptive changes that decrease its SDS elutability, displaceability by plasma, polyclonal antifibrinogen binding, and ability to support platelet adhesion (summarized in Chinn et al. J. Biomed. Mater. Res. 26, 757 (1992)). In order to better understand the nature of the changes in adsorbed fibrinogen, the binding of ten different monoclonal antifibrinogen molecules to fibrinogen adsorbed from plasma to Biomer and several other surfaces has been measured after increasing residence time in buffer. Three of the monoclonal antibodies used bind to sequences that have been implicated in platelet binding to fibrinogen. One of these (M1) binds to the C-terminal region of the gamma chain (402-411), another (R1) binds to the N-terminal region of the A alpha chain containing an RGDF sequence (95-98), and the third (R2) binds to the C-terminal region of the A alpha chain containing an RGDS sequence (572-575). Two other antibodies (P1 and K4) also bind to the C-terminal region of the gamma chain (373-385 and 392-406, respectively). Five other antibodies that bind to other regions in fibrinogen were also used. Two of the antibodies (K4 and P1) are also known to be sensitive to conformational changes in the fibrinogen molecule. The binding of the various antibodies changed with residence time in ways that were highly dependent on the particular antibody. The binding of some antibodies was very stable with respect to residence time, others rose with time, some declined with residence time and one appears to pass through a maximum. However, none of the changes in antibody binding were nearly as fast as has been observed for the changes in platelet binding reported previously. Binding to the platelet binding region near the gamma chain C-terminal region either did not change with residence time (M1), increased with residence time (K4), or else decreased more slowly than observed for platelets (P1). Binding of the antibodies to the RGD sequences near the N-terminus of the A alpha chain (95-98) was very low initially but increased with residence time, while the binding to the RGD sequence near the C-terminus of the A alpha chain (572-575) increased slightly at short residence times but then declined substantially after longer residence times. Thus, the changes in the expression of the putative platelet binding domains do not correlate with the declines in platelet binding to plasma preadsorbed Biomer.(ABSTRACT TRUNCATED AT 400 WORDS)
authors.affiliation: Department of Chemical Engineering and Bioengineering, University of Washington, Seattle 98195.
authors.forename: T A
authors.initials: TA
authors.lastname: Horbett
authors.forename: K R
authors.initials: KR
authors.lastname: Lew
mesh_terms: D000327:Adsorption; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D000937:Antigen-Antibody Reactions; D001672:Biocompatible Materials; D002021:Buffers; D004797:Enzyme-Linked Immunosorbent Assay; D005340:Fibrinogen / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009842:Oligopeptides; D010446:Peptide Fragments / Q000276:immunology / Q000378:metabolism; D010973:Platelet Adhesiveness; D011140:Polyurethanes; D015203:Reproducibility of Results; D013997:Time Factors
chemical_list: D000911:Antibodies, Monoclonal; D001672:Biocompatible Materials; D002021:Buffers; D009842:Oligopeptides; D010446:Peptide Fragments; D011140:Polyurethanes; C031892:biomer; C047981:arginyl-glycyl-aspartic acid; D005340:Fibrinogen