Detection of factor X activation in humans.
A sensitive radioimmunoassay (RIA) for the fragment that is liberated from factor X when this zymogen is activated by factor VII/VIIa-tissue factor or factor IXa was developed. Antisera were raised in rabbits to a synthetic 15 amino acid peptide containing the COOH-terminal sequence of the activation fragment coupled to bovine serum albumin with glutaraldehyde. The reactivity of the antibody population obtained toward the factor X zymogen was negligible (less than 1/36,000 that of the activation peptide on a molar basis). However, because other plasma constituents contributed to a nonspecific basal signal in the RIA, a procedure by which the peptide could be reproducibly extracted from plasma was developed. The mean level of this species in normal individuals younger than the age of 40 was 66.4 pmol/L, and elevations up to 550 pmol/L were observed in patients with evidence of disseminated intravascular coagulation. The validity of these measurements of factor X activation is supported by the fact that the RIA signal migrates on reverse-phase high pressure liquid chromatography in a manner identical to that of the native peptide and can be quantitatively recovered. The mean concentration of the activation fragment was markedly decreased to 25.7 pmol/L in patients with hereditary factor VII deficiency (P = .0001 v normal controls), whereas the mean level in subjects with factor VIII deficiency was 61.1 pmol/L (P greater than .1 v normal controls). These data indicate that the basal (ie, in the absence of thrombosis or provocative stimuli) levels of FXP under in vivo conditions result mainly from the activity of the extrinsic pathway.
Bauer
K A
KA
Charles A. Dana Research Institute, Beth Israel Hospital, Boston, MA 02215.
Kass
B L
BL
ten Cate
H
H
Bednarek
M A
MA
Hawiger
J J
JJ
Rosenberg
R D
RD
D000818:Animals; D000925:Anticoagulants / Q000494:pharmacology; D001777:Blood Coagulation; D004211:Disseminated Intravascular Coagulation / Q000097:blood; D004285:Dogs; D005168:Factor VII Deficiency / Q000097:blood; D005170:Factor X / Q000378:metabolism*; D015951:Factor Xa / Q000378:metabolism; D006467:Hemophilia A / Q000097:blood; D002836:Hemophilia B / Q000097:blood; D006801:Humans; D010446:Peptide Fragments / Q000032:analysis; D011863:Radioimmunoassay
D000925:Anticoagulants; D010446:Peptide Fragments; D005170:Factor X; D015951:Factor Xa