Fischer R R, Lucas E M, Pereira A M, Roisenberg I
Departamento de Genética, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brasil.
Braz J Med Biol Res. 1996 Dec;29(12):1641-4.
A simple method for the preparation of rabbit antiserum against human von Willebrand factor (vWF) from commercial lyophilized factor VIII concentrate is described. vWF antigen (vWFAg)-like protein was obtained by gel filtration of the concentrate on Sepharose 4B. A combination of measurements of protein content by absorbance at 280 nm, and of vWFAg by electroimmunoassay using a commercial antibody, provided the data needed to select the Sepharose-filtered fractions with the highest concentrations of vWFAg-like protein. The immunization scheme used induced high antibody titers from the 45th to the 126th day after the first immunization. The resulting antiserum showed a performance similar to that of a commercial preparation in terms of vWFAg determination by electroimmunoassay and two-dimensional crossed-immunoelectrophoresis.
描述了一种从市售冻干因子VIII浓缩物制备兔抗人血管性血友病因子（vWF）抗血清的简单方法。通过在琼脂糖4B上对浓缩物进行凝胶过滤获得vWF抗原（vWFAg）样蛋白。通过在280nm处测量吸光度来测定蛋白质含量，并使用市售抗体通过免疫电泳测定vWFAg，这些测量相结合提供了选择具有最高浓度vWFAg样蛋白的琼脂糖过滤级分所需的数据。所采用的免疫方案在首次免疫后第45天至第126天诱导出高抗体滴度。通过免疫电泳和二维交叉免疫电泳测定vWFAg，所得抗血清的性能与市售制剂相似。
