-transparent.png){kind=logo}
Carboxypeptidase U (EC 3.4.17.20, CPU, TAFIa) is a novel determinant of the fibrinolytic rate. It circulates as an inactive zymogen, procarboxypeptidase U, which becomes active during the process of coagulation. We developed a high throughput method on microtiter plates for the determination of the procarboxypeptidase U concentration in human plasma samples. Following activation of procarboxypeptidase U by thrombin-thrombomodulin, the resulting enzyme activity cleaves p-OH-Hip-Arg and the generated p-OH-hippuric acid is converted by hippuricase to p-hydroxybenzoic acid and glycine. Finally, oxidative coupling of p-hydroxybenzoic acid with 4-aminoantipyrine by NaIO4 forms the quinoneimine dye. The absorbance of the latter dye is determined at 506 nm in a microtiter plate reader. A mean value of 620 U/l was found, with a CV of 3.0% within-run and 4.3% between-run. The assay showed a good correlation with the activities observed using a HPLC assay as reference method (n = 25, r = 0.979). The presented method enables the routine analysis of large sample pools in clinical setting.
K A Schatteman, and
F J Goossens, and
J Leurs, and
Y Kasahara, and
S S Scharpé, and
D F Hendriks
October 2005,
Journal of thrombosis and haemostasis : JTH,
K A Schatteman, and
F J Goossens, and
J Leurs, and
Y Kasahara, and
S S Scharpé, and
D F Hendriks
July 2003,
Journal of thrombosis and haemostasis : JTH,
K A Schatteman, and
F J Goossens, and
J Leurs, and
Y Kasahara, and
S S Scharpé, and
D F Hendriks
March 2001,
Thrombosis research,
K A Schatteman, and
F J Goossens, and
J Leurs, and
Y Kasahara, and
S S Scharpé, and
D F Hendriks
January 2001,
Thrombosis and haemostasis,
K A Schatteman, and
F J Goossens, and
J Leurs, and
Y Kasahara, and
S S Scharpé, and
D F Hendriks
January 2004,
Seminars in hematology,
K A Schatteman, and
F J Goossens, and
J Leurs, and
Y Kasahara, and
S S Scharpé, and
D F Hendriks
January 2010,
Analytical biochemistry,
K A Schatteman, and
F J Goossens, and
J Leurs, and
Y Kasahara, and
S S Scharpé, and
D F Hendriks
January 2006,
Clinical chemistry,
K A Schatteman, and
F J Goossens, and
J Leurs, and
Y Kasahara, and
S S Scharpé, and
D F Hendriks
September 2004,
Clinica chimica acta; international journal of clinical chemistry,
K A Schatteman, and
F J Goossens, and
J Leurs, and
Y Kasahara, and
S S Scharpé, and
D F Hendriks
September 2006,
Analytical biochemistry,
K A Schatteman, and
F J Goossens, and
J Leurs, and
Y Kasahara, and
S S Scharpé, and
D F Hendriks
September 2002,
Clinical chemistry,
