Cooper H A, Lee D, Lamb M A, Wagner R H
Br J Haematol. 1980 Jan;44(1):149-60. doi: 10.1111/j.1365-2141.1980.tb01192.x.
An antibody was raised in rabbits to the small active fragment of human factor VIII, obtained by Ca2+ dissociation of a human factor VIII preparation made from a multidonor plasma pool. After absorption, the antibody neutralized the factor VIII coagulant activity of normal human plasma, but did not precipitate with any plasma or plasma fractions or neutralize von Willebrand factor (vwf) activity as measured by ristocetin aggregation of fixed washed platelets. Immune beads were prepared by CNBr binding of the partially purified rabbit antibody to 1% agarose beads. Non-immune beads were prepared with IgG fractions obtained from the rabbits before immunization and used throughout as a control. The amount of factor VIII coagulant activity (VII:C) removed from plasma by immune beads was time-dependent and proportional to the amount of beads used, but all of the VIII:C could not be readily removed. Removal of VIII:C by immune beads parallelled removal of factor VIII:antigen, but less VWF activity was removed. Immune beads could be blocked or saturated by treatment with large amounts of normal plasma, but not by von Willebrand disease plasma and only by some haemophilic plasma.
用人因子 VIII 的小活性片段免疫家兔制备抗体，该小活性片段是通过从多供体血浆库制备的人因子 VIII 制剂经 Ca2+ 解离获得的。吸收后，该抗体可中和正常人血浆中的因子 VIII 凝血活性，但不与任何血浆或血浆组分沉淀，也不中和用固定洗涤血小板的瑞斯托霉素聚集法测定的血管性血友病因子（vwf）活性。通过将部分纯化的兔抗体与 1% 琼脂糖珠进行溴化氰结合制备免疫珠。用免疫前从兔获得的 IgG 组分制备非免疫珠，并始终用作对照。免疫珠从血浆中去除的因子 VIII 凝血活性（VII:C）是时间依赖性的，且与所用珠的量成正比，但并非所有 VIII:C 都能轻易去除。免疫珠去除 VIII:C 与去除因子 VIII:抗原平行，但去除的 VWF 活性较少。用大量正常人血浆处理可封闭或饱和免疫珠，但血管性血友病血浆不能，只有一些血友病血浆能做到。
