ABNORMAL FACTOR-VIII HIROSHIMA - DEFECT IN CRUCIAL PROTEOLYTIC CLEAVAGE BY THROMBIN AT ARG(1689) DETECTED BY A NOVEL ELISA
ABNORMAL FACTOR-VIII HIROSHIMA - DEFECT IN CRUCIAL PROTEOLYTIC CLEAVAGE BY THROMBIN AT ARG(1689) DETECTED BY A NOVEL ELISA
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DOI：
10.1111/j.1365-2141.1994.tb03259.x
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发表时间：
1994-01-01
影响因子：
6.5
通讯作者：
YOSHIOKA, A
中科院分区：
文献类型：
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作者：
KAMISUE, S;SHIMA, M;YOSHIOKA, A
我们建立了检测凝血酶在Arg（1689）处切割FVIII轻链的ELISA。该方法使用识别C2结构域中氨基酸残基2248-2312的包被同种抗体，以及识别轻链氨基末端区域中残基1675-1684的第二单克隆抗体NMC-VIII/10。在用不同浓度的凝血酶处理血浆后测量FVIII抗原（FVIII：Ag）。正常血浆的FVIII：Ag以剂量依赖性方式被凝血酶降低，在1.0 U/ml酶的存在下降至28%。达到50%降低的凝血酶浓度（IC 50）约为1.0 U/ml。分析了4例血友病A阳性（A（+））和2例血友病A减少（AR）患者的血浆。所有患者的IC 50均大于1.0 U/ml，表明凝血因子VIII轻链的凝血酶切割存在缺陷。一份血友病A+血浆对凝血酶没有反应，
We have established an ELISA for detecting thrombin cleavage of the FVIII light chain at Arg(1689), The method used a coating alloantibody which recognized amino acid residues 2248-2312 in the C2 domain, together with a second monoclonal antibody, NMC-VIII/10, which recognized residues 1675-1684 in the amino-terminal region of the light chain. FVIII antigen (FVIII:Ag) was measured after treatment of plasma with various concentrations of thrombin. The FVIII:Ag of normal plasma was reduced in a dose-dependent manner by the thrombin, falling to 28% in the presence of 1.0 U/ml enzyme. The concentration of thrombin that achieved 50% reduction (IC50) was approximately 1.0 U/ml. The plasma of four haemophilia A positive (A(+)) and two haemophilia A reduced (AR) patients were analysed. The IC50 of all patients was more than 1.0 U/ml, indicating that thrombin cleavage of the FVIII light chain was defective. One haemophilia A+ plasma did not respond to thrombin in