A soluble fibrin standard: comparable dose-response with immunologic and functional assays.
A soluble fibrin (SF) preparation has been developed as a potential standard by the Scientific and Standardization Committee for use in assays evaluating in vitro preparations and patient plasma samples. The SF standard was prepared by reaction of factor XIII-free fibrinogen with thrombin, followed by neutralization with hirudin and solubilization of the fibrin in acetic acid. As characterized by SDS-PAGE, the polypeptide chain structure shows the anticipated loss of fibrinopeptides and lack of gamma or alpha chain crosslinking. The standard was added to pooled normal plasma at concentrations from 12.5 microg/ml to 340 microg/ml and tested with four commercially available assays based on immunologic reactions using ELISA or latex agglutination or on t-PA cofactor activity for plasminogen to plasmin conversion. Absolute "soluble fibrin" concentrations were calculated using the manufacturers' calibrators and showed distinct dose-response relationships for each assay. Expression of the results following log-transformation produced a series of parallel lines, indicating that this SF preparation can serve as a standard, effectively normalizing the disparate proprietary internal calibrators currently used for each assay.
McCarron
B I
BI
Vascular Medicine Unit, Department of Medicine, University of Rochester School of Medicine and Dentistry, New York, USA.
Marder
V J
VJ
Kanouse
J J
JJ
Francis
C W
CW
D001681:Biological Assay; D004305:Dose-Response Relationship, Drug; D005337:Fibrin / Q000032:analysis / Q000592:standards*; D006801:Humans; D007118:Immunoassay; D019934:International Normalized Ratio
D005337:Fibrin