Biochemical and functional characterization of recombinant von Willebrand factor produced on a large scale

1997 ◽  
Vol 53 (12) ◽  
pp. 943 ◽  
Author(s):  
B. E. Fischer ◽  
U. Schlokat ◽  
M. Reiter ◽  
W. Mundt ◽  
F. Dorner
Keyword(s):  
Von Willebrand Factor ◽  
Large Scale ◽  
Functional Characterization ◽  
Von Willebrand ◽  
Willebrand Factor

Cloning, expression, and functional characterization of the von Willebrand factor–cleaving protease (ADAMTS13)

Blood ◽  
2002 ◽  
Vol 100 (10) ◽  
pp. 3626-3632 ◽  
Author(s):  
Barbara Plaimauer ◽  
Klaus Zimmermann ◽  
Dirk Völkel ◽  
Gerhard Antoine ◽  
Randolf Kerschbaumer ◽  
...  
Keyword(s):  
Von Willebrand Factor ◽  
Functional Activity ◽  
Functional Characterization ◽  
Eukaryotic Expression ◽  
Gene Encoding ◽  
Hek 293 ◽  
293 Cells ◽  
Von Willebrand ◽  
Willebrand Factor

Deficient von Willebrand factor (VWF) degradation has been associated with thrombotic thrombocytopenic purpura (TTP). In hereditary TTP, the specific VWF-cleaving protease (VWF-cp) is absent or functionally defective, whereas in the nonfamilial, acquired form of TTP, an autoantibody inhibiting VWF-cp activity is found transiently in most patients. The gene encoding for VWF-cp has recently been identified as a member of the metalloprotease family and designatedADAMTS13, but the functional activity of the ADAMTS13 gene product has not been verified. To establish the functional activity of recombinant VWF-cp, we cloned the complete cDNA sequence in a eukaryotic expression vector and transiently expressed the encoded recombinant ADAMTS13 in HEK 293 cells. The expressed protein degraded VWF multimers and proteolytically cleaved VWF to the same fragments as those generated by plasma VWF-cp. Furthermore, recombinant ADAMTS13-mediated degradation of VWF multimers was entirely inhibited in the presence of plasma from a patient with acquired TTP. These data show that ADAMTS13 is responsible for the physiologic proteolytic degradation of VWF multimers.

Identification and Functional Characterization of von Willebrand Disease

2009 ◽  
Vol 02 ◽  
pp. 16
Author(s):  
Emmanuel J Favaloro ◽  
Keyword(s):  
Von Willebrand Factor ◽  
Classification Scheme ◽  
Binding Assay ◽  
Functional Characterization ◽  
Von Willebrand Disease ◽  
Collagen Binding ◽  
Diagnostic Approaches ◽  
Von Willebrand ◽  
Willebrand Factor

The identification and functional characterization of von Willebrand disease (VWD) is challenging due to clinical uncertainty and limitations in test processes and panels used by laboratories, and because the classification scheme does not always permit unequivocal assignment of subtype. This article reviews contemporary alternatives to classic diagnostic approaches, including the incorporation of extended core test panels inclusive of the collagen-binding assay, and the potential for desmopressin (DDAVP) challenges not only to provide therapeutic information but also to assist the better characterization of individuals with defects or deficiencies in von Willebrand factor (VWF). Supplementary assays such as the PFA-100® and the VWF propeptide assay following DDAVP challenge are also worth considering.

Functional characterization of bovine von Willebrand factor gene promoter in bovine endothelial cells demonstrates species-specific properties

Gene ◽  
1997 ◽  
Vol 198 (1-2) ◽  
pp. 127-134 ◽  
Author(s):  
Nathalie Janel ◽  
Anne-Marie Dosne ◽  
Bernadette Obert ◽  
Dominique Meyer ◽  
Danièle Kerbiriou-Nabias
Keyword(s):  
Endothelial Cells ◽  
Von Willebrand Factor ◽  
Functional Characterization ◽  
Gene Promoter ◽  
Factor Gene ◽  
Bovine Endothelial Cells ◽  
Species Specific ◽  
Von Willebrand ◽  
Willebrand Factor

Characterization of Partial Gene Deletions in Type III von Willebrand Disease with Alloantibody Inhibitors

1994 ◽  
Vol 72 (02) ◽  
pp. 180-185 ◽  
Author(s):  
David J Mancuso ◽  
Elodee A Tuley ◽  
Ricardo Castillo ◽  
Norma de Bosch ◽  
Pler M Mannucci ◽  
...  
Keyword(s):  
Von Willebrand Factor ◽  
Von Willebrand Disease ◽  
Pcr Analysis ◽  
Gene Deletions ◽  
Factor Gene ◽  
Type Iii ◽  
Large Deletions ◽  
Von Willebrand ◽  
Willebrand Factor

Summaryvon Willebrand factor gene deletions were characterized in four patients with severe type III von Willebrand disease and alloantibodies to von Willebrand factor. A PCR-based strategy was used to characterize the boundaries of the deletions. Identical 30 kb von Willebrand factor gene deletions which include exons 33 through 38 were identified in two siblings of one family by this method. A small 5 base pair insertion (CCTGG) was sequenced at the deletion breakpoint. PCR analysis was used to detect the deletion in three generations of the family, including two family members who are heterozygous for the deletion. In a second family, two type III vWD patients, who are distant cousins, share an -56 kb deletion of exons 22 through 43. The identification and characterization of large vWF gene deletions in these type III vWD patients provides further support for the association between large deletions in both von Willebrand factor alleles and the development of inhibitory alloantibodies.

scholarly journals Substructure of human von Willebrand factor. Proteolysis by V8 and characterization of two functional domains.

1986 ◽  
Vol 261 (33) ◽  
pp. 15679-15689 ◽  
Author(s):  
L J Fretto ◽  
W E Fowler ◽  
D R McCaslin ◽  
H P Erickson ◽  
P A McKee
Keyword(s):  
Von Willebrand Factor ◽  
Functional Domains ◽  
Von Willebrand ◽  
Willebrand Factor
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