Production and characterization of monoclonal antibodies against von Willebrand factor (vWF)
Production and characterization of monoclonal antibodies against von Willebrand factor (vWF)
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抗血管性血友病因子 (vWF) 单克隆抗体的生产和表征
DOI：
10.2491/jjsth1970.16.205
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发表时间：
1985
期刊：
影响因子：
--
通讯作者：
H. Fukui
中科院分区：
文献类型：
--
作者：
M. Shima;J. Morimoto;S. Imai;Y. Tsubura;A. Yoshioka;H. Fukui
用FVIII/vWF部分免疫BALB/C小鼠，并将脾细胞与NS-1小鼠骨髓瘤细胞融合，制备抗血管性血友病因子（vWF）单克隆抗体。采用酶联免疫吸附试验（ELISA）检测产生抗体的杂交瘤。该方法获得的5个克隆单克隆抗体均属于IgGI类，通过玻璃微珠抑制血小板滞留（Bowie等）。抑制作用以3B11最为显著。ELISA检测的vWF: Ag抗体滴度范围为1 × 105 ~ 1 × 107。3B11抑制里斯托司汀诱导的血小板聚集和里斯托司汀辅助因子达1 × 104。单克隆抗体免疫吸附层析法分别从FVIII/vWF组分中洗脱VIII蛋白和vWF蛋白。建立了一种新的ELISA方法，利用各单克隆抗体定量测定vWF: Ag。下限是0。001 u /毫升。采用单克隆抗体亲和素-生物素系统免疫过氧化物酶染色法检测组织中vWF: Ag的表达。新生儿脐带内皮细胞、水洗血小板和巨核细胞染色阳性。
Monoclonal antibodies against von Willebrand factor (vWF) were produced by immunization of BALB/C mice with FVIII/vWF fraction and fusion of spleen cells with NS-1 murine myeloma cells. Antibody producing hybridomas were detected by an enzyme linked immunosorbent assay (ELISA). Five cloned monoclonal antibodies obtained in this method were all belonged to IgGI class, and inhibited platelet retention by glass beads (Bowie et al.). The inhibitory action was most remarkable in 3B11. The antibody titer against vWF: Ag ranged from 1 X 105 to 1 X 107 by an ELISA. 3B11 inhibited ristocetin induced platelet aggregation and ristocetin cofactor up to 1 X 104. VIII protein and vWF protein were separately eluted from FVIII/vWF fraction by immunoadsorbent chromatography using monoclonal antibody. A new ELISA method was developed using each monoclonal antibody to quantitate vWF: Ag. The lower limit was 0. 001 u/ml. In order to detect vWF: Ag in tissues immunoperoxidase staining by avidin-biotin system using monoclonal antibody was performed. Positive staining was obtained in the endothelial cells of the neonatal umbilical cord, washed platelets and megakaryocytes.