Localization of a factor VIII-inhibiting antibody epitope to a region between residues 338 and 362 of factor VIII heavy chain.
Localization of a factor VIII-inhibiting antibody epitope to a region between residues 338 and 362 of factor VIII heavy chain.
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因子 VIII 抑制抗体表位定位于因子 VIII 重链残基 338 和 362 之间的区域。
DOI：
10.1073/pnas.85.9.3165
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发表时间：
1988
影响因子：
11.1
通讯作者：
Stafford,DW
中科院分区：
文献类型：
--
作者：
Ware,J;Toomey,JR;Stafford,DW
我们使用重组DNA表位文库来定位因子VIII （FVIII）单克隆抗体的结合区，该抗体可以中和凝血剂活性。该抗体C5先前已被描述，并已被证明具有每毫克纯化免疫球蛋白1488贝塞斯达单位的FVIII中和效力。利用短而随机的FVIII cDNA片段构建重组DNA表位文库，并用C5进行免疫筛选，以鉴定表达抗原决定因子的噬菌体。免疫反应性噬菌体的分离和鉴定将C5表位限制在9个不同克隆的重叠或共享的DNA序列上，并且与成熟的FVIII肽的氨基酸残基338-362相对应。确定的表位位于90 kda FVIII重链亚基氨基末端的活化蛋白C裂解位点（Arg-336）和凝血酶裂解位点（Arg-372）之间。在两个关键切割位点之间鉴定了抑制抗FVIII抗体的表位，表明该氨基酸序列在调节FVIII凝血活性中起作用。
We have used a recombinant DNA epitope library to localize the binding region of a factor VIII (FVIII) monoclonal antibody that neutralizes coagulant activity. The antibody, C5, has previously been described and has been shown to have a FVIII neutralizing potency of 1488 Bethesda units per mg of purified immunoglobulin. A recombinant DNA epitope library was constructed from short, random FVIII cDNA fragments and immunologically screened with C5 to identify bacteriophage expressing the antigenic determinant. The isolation and characterization of immunoreactive bacteriophage restricted the C5 epitope to the overlapping or shared DNA sequence of nine different clones and corresponded to amino acid residues 338-362 of the mature FVIII peptide. The defined epitope is between the proposed activated protein C cleavage site (Arg-336) and thrombin cleavage site (Arg-372) on the amino-terminal 90-kDa FVIII heavy-chain subunit. The identification of the epitope of an inhibiting anti-FVIII antibody between two critical cleavage sites suggests that this amino acid sequence plays a role in regulating FVIII coagulant activity.