Study on functinal domains of von Willebrand (vW) factor in patients with vW disease.

1 .血管性血友病因子（vWf）的功能域分析及vWf单克隆抗体的制备和鉴定。用DEAE和肝素柱纯化因子viii结合域：vwf -纤溶酶或胰蛋白酶消化片段p-34（氨基酸残基1-298(273)）。4种不同的针对p-34的单克隆抗体由D.Meyer， INSERM 143法国，与我合作。现在，我要描述这些抗体b。血小板糖蛋白Ib（GPIb）结合域：两种不同的单克隆抗体（NMC/vW4）。对抑制vWf与GPIb结合的两种诱导剂进行了表征，并发现了两种诱导剂（抗菌素里斯托韦特素和毒液因子botrocetin）对GPIb与vWf结合的不同功能位点。已知Aurin三羧酸（ATA）抑制利斯托司汀诱导的血小板聚集，但对其他血小板聚集物无抑制作用。进一步研究了其消除人vWf-血小板相互作用的能力，发现ATA通过干扰vWf而不干扰血小板来阻断vWf/GPIb通路，是预防早期血栓形成的潜在工具。人胶原结合结构域：发现1个抑制vWf-人iii型原纤维胶原结合的单抗（NMC/vW3），其表位位于vWf亚基。即表位定位于氨基酸残基911和1365.II之间。血管性血友病患者血浆血管性血友病因子分析。我研究了几种正常和几种血管性血友病血浆vWf功能测定和vWf蛋白分析的建立。建立了使用微量血浆的vwf -人胶原蛋白结合试验。ⅱ型vWd患者胶原结合异常。建立了用微量样品分析等离子体vWf中vWf亚基的方法。现在，我们将对几种vWd类型中的等离子体vWf进行检查。

I. The analysis of functional domains of von Willebrand factor(vWf) and the production and characterization of monoclonal antibodies (MAbs) against vWf.A. the factorVIII-binding domain: vWf-plasmin or trypsin digested fragment p-34(amino acid residue 1-298(273)) was purified using DEAE and Heparin column. 4 different MAbs against p-34 were produced by D.Meyer, INSERM 143 France, collaborated with me. Now, I'm going to characterize these antibodiesB. Platelet glycoprotein Ib(GPIb)-binding domain: Two different MAbs (NMC/vW4 from our lab. 322 from france) against vWf which inhibit the binding of vWf to GPIb were characterized, and I discovered the different functional site of GPIb-binding of vWf by two inducers, such as antibiotics ristocetin and venom factor botrocetin. and Aurin tricarboxylic acid (ATA) is known to inhibit ristocetin-induced platelet aggregation but not the other platelet aggregaters. Its capacity to abolish human vWf-platelet interaction was further in vestigated and was found that ATA which blocks the vWf/GPIb pathway by interfering with vWf and not with platelet, is a potential tool in prevention the early stages of thrombosisC. Human collagen-binding domain: One MAb(NMC/vW3) which inhibit the vWf-human typeIII fibrilar collagen binding was discovered and was shown the epitope in vWf subunit. That is, The epitope was localized between the amino acid residue 911 and 1365.II. The analysis of plasma von Willebrand factor in von Willebrand disease. I studied on the establishment of several vWf function assays and vWf protein analysis of plasma vWf in normal and several types of von Willebrand disease.A. The vWf-human collagen binding assay, using a minute plasma was established. The abnormality of the collagen- binding in type II vWd was discovered.B. The analytical method of vWf subunit in plasma vWf using a minute sample was established. Now the plasma vWf in several types of vWd is going to examine.