geoscience.net logo
+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

Estimation of the von Willebrand factor-cleaving protease in plasma using monoclonal antibodies to vWF


, : Estimation of the von Willebrand factor-cleaving protease in plasma using monoclonal antibodies to vWF. Thrombosis and Haemostasis 82(5): 1382-1385

A protease present in plasma cleaves von Willebrand factor (vWF) at the peptide bond 842Tyr-843Met of the mature subunit. To quantify this vWF-cleaving protease activity in plasma we have developed a simple method based on the estimation by IRMA of the degradation of a constant amount of wild type recombinant vWF used as substrate, by serial dilutions of test plasma used as protease provider. vWFAg was estimated by two-site IRMA using as first coating antibody a monoclonal antibody (MoAb) whose epitope is localized on the C-terminal side of the cleavage site, and as second labeled antibody a pool of MoAbs specific for the N-terminal side. Because the proteolytic process leads to the progressive separation of the C- and N-terminal portions of the vWF subunit such an IRMA also shows a progressive apparent loss of vWFAg. In contrast, the levels of vWFAg estimated after proteolysis by regular IRMA remained essentially constant. Results obtained with this new method were compared with the analysis by SDS-agarose gel electrophoresis of the multimeric pattern of proteolyzed WT-rvWF and no significant difference was noted testing a series of 28 plasmas. As compared with normal pooled plasma, 14 normal individuals and 13 patients with various types of vWD had normal levels of protease activity (44-178%) by both methods. The validity of the method was confirmed by showing a lack of detectable protease activity in a patient with chronic relapsing thrombotic thrombocytopenic purpura. In conclusion our method appears as a useful tool for the quantification of the vWF-cleaving protease activity in plasma. Its sensitivity and specificity are similar to those of SDS-gel electrophoresis. However, this new IRMA has the major advantages of being much simpler and faster, and open to most research laboratories in the field.

US$29.90

PMID: 10595622


Other references

Huysmans, S.; E.G.azaly, G.; Smets, E., 2000: Orbicules: still a well-hidden secret of the anther. Orbicules (or Ubisch bodies) are small structures of sporopollenin (<1 to 15 micro m). This chapter deals mainly with the distribution of orbicules in the angiosperms and their possible function(s). To find possible clues in the systematic dist...

Saunders, L.G., 1958: The pollination of cacao. The known pollinators of cacao are shown to belong to the subgenera Proforcipomyia and Thyridomyia of the genus Forcipomyia. Characters are presented for the recognition of larvae, pupae, and adults of members of these groups. The breeding grounds...

Chen, B-Yu.; Kuo, C-Ching.; Huang, Y-Shao.; Lu, S-Tung.; Liang, F-Cheng.; Jiang, D-Hua., 2015: Novel highly selective and reversible chemosensors based on dual-ratiometric fluorescent electrospun nanofibers with pH- and Fe(3+)-modulated multicolor fluorescence emission. Novel dual-ratiometric fluorescent electrospun (ES) nanofibers featuring high sensitivity for pH and ferric ion (Fe(3+)) were prepared using binary blends of poly(2-hydroxyethyl methacrylate-co-N-methylolacrylamide-co-nitrobenzoxadiazolyl derivati...

Hunt T.W.; Monaco T.J.; Sheets T.J., 1980: Residue and efficacy studies of alachlor on cabbage brassica oleracea var capitata f alba cultivar market prize and soil. Newly transplanted cabbage (B. oleracea L. var. capitata L. f. alba DC. cv. Market Prize) was treated with alachlor [2 chloro-2',6'-diethyl-N-(methoxymethyl)acetamide] at rates of 0, 3.4 and 6.7 kg/ha in 4 tests of 2 locations during 197...

Kertesz, V.; Van Berkel, G.J., 2008: Improved desorption electrospray ionization mass spectrometry performance using edge sampling and a rotational sample stage. The position of the surface to be analyzed relative to the sampling orifice or capillary into the mass spectrometer has been known to dramatically affect the observed signal levels in desorption electrospray ionization mass spectrometry (DESI-MS)....

Rinderknecht, Andres, 2000: La presencia de osteodermos en las extremidades posteriores de Glyptodon clavipes Owen, 1839 (Mammalia: Cingulata). Hind limb osteoderms of an individual assigned to Glyptodon clavipes Owen, from the Pleistocene of Uruguay are described. A great number of these bones were observed in the foot, as well as in the calf and thigh. These elements had been cited for...

E.H.nnawy, S.; Eid, S.A.; Hussein, K.R.F.; Tarrad, A.F., 1981: Effect of water levels on the biosynthesis of lipids and fatty acids in sunflower seeds. Oil contents of seeds of sunflower cv. Meiak and Giza-1 increased with seed maturity, but decreased with decreased irrigation. Highest amounts of oils accumulated between 14 and 28 days after flowering. Palmitic, oleic, linoleic and arachidic acid...

Gérard, H.C.; Köhler, L.; Branigan, P.J.; Zeidler, H.; Schumacher, H.R.; Hudson, A.P., 1998: Viability and gene expression in Chlamydia trachomatis during persistent infection of cultured human monocytes. The principal host cell for persistently infecting synovial Chlamydia trachomatis is the macrophage. During infection of human monocytes/macrophages in culture this bacterium displays aberrant morphology and produces no new elementary bodies, refl...

Flanagan, F.; Barrett, D.R., 1993: Sandalwood nuts as food. Sandalwood (Santalum spicatum) fruits were collected from trees growing near Narrogin, Western Australia. The fibrous exocarp and the hard endocarp were removed to reveal the kernel (endosperm and embryo). The nutritive value of sandalwood nuts wa...

Debaene, B.; Meistelman, C.; Beaussier, M.; Lienhart, A., 1994: Visual estimation of train-of-four responses at the orbicularis oculi and posttetanic count at the adductor pollicis during intense neuromuscular block. Posttetanic count (PTC) predicts the time to return of train-of-four (TOF) responses at the adductor pollicis (AP) muscle. The duration of neuromuscular block at the orbicularis oculi (OO) is shorter than at the AR The aim of this study was to ass...