[Clinical application of a method for evaluating von Willebrand factor cleaving protease activity].
OBJECTIVE
To establish a method for evaluating the activity of von Willebrand factor cleaving protease (vWF-cp) and evaluate its clinical application.


METHODS
Purified von Willebrand factor polymer was isolated by gel filtration from human fresh-frozen plasma as the enzyme substrate. SDS-PAGE, Western blotting, and luminographic detection were used to evaluate vWF-cp activity of 60 healthy adults, 28 patients with cerebral infarction (CI) and 7 with thrombotic thrombocytopenic purpura (TTP).


RESULTS
In the subjects involved, the method for evaluating vWF-cp activity had intra- and inter-batch coefficient of variation(CV) of 4.81% (n=8) and 8.63% (n=6), respectively. According to this method, the plasma vWF-cp activity in the 60 healthy adults was significantly higher than that in the CI patients [(86.53-/+17.49)% vs (77.15-/+16.72)%, P<0.05]. In TTP patients before plasma replacement, the vWF-cp activity was (9.06-/+7.17)% and increased significantly to (47.00-/+6.27)% 24 h after plasma replacement, respectively, but still significantly lower than that of healthy adults (P<0.01), whereas in the convalescent stage, the activity approached the normal level [(83.18-/+8.83)%, P>0.05].


CONCLUSIONS
According to the described method, which allows accurate vWF-cp activity measurement with good sensitivity, specificity and reproducibility, vWF-cp activity is lower in CI patients and even more so in TTP patients than that of healthy adults. Plasma replacement can effectively increase the vWF-cp activity in TTP patients.
Lin
Fa-Quan
FQ
Clinical Medical Experiment Center, First Affiliated Hospital, Guangxi Medical University, Nanning 530021, China. fqlin1998@163.com
Li
Ming-Fen
MF
Liu
Qian
Q
Li
Zhao-Yan
ZY
D051722:ADAM Proteins / Q000097:blood / Q000378:metabolism*; D000071120:ADAMTS13 Protein; D000328:Adult; D000818:Animals; D002544:Cerebral Infarction / Q000097:blood / Q000201:enzymology; D057075:Enzyme Assays / Q000379:methods*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D011697:Purpura, Thrombotic Thrombocytopenic / Q000097:blood / Q000201:enzymology; D015203:Reproducibility of Results; D013379:Substrate Specificity
D051722:ADAM Proteins; D000071120:ADAMTS13 Protein; C099604:ADAMTS13 protein, human