Berenson R J, Bensinger W I, Kalamasz D, Martin P
Blood. 1986 Feb;67(2):509-15.
Biotinylated antibodies and an avidin-Sepharose 6MB column were utilized in a novel approach to deplete selected cell populations from human bone marrow. Fluorescein-labeled Daudi lymphoblasts were mixed with bone marrow mononuclear cells in a model system, and removal of Daudi cells was quantitatively assessed with an inverted fluorescent microscope. Treatment using the biotinylated monoclonal antibody 2H7 reactive with Bp32 antigen (expressed on Daudi cells) followed by passage over avidin-Sepharose produced greater than two logs of Daudi cell removal from bone marrow. An alternative method was tested by treating cells successively with nonbiotinylated monoclonal antibody and biotinylated goat antimouse immunoglobulin followed by passage over avidin-Sepharose. Up to three logs of Daudi cells could be eliminated from bone marrow with quantitative recovery of hematopoietic progenitors. The use of biotinylated goat antimouse immunoglobulin eliminates the need to prepare a biotin conjugate of each individual monoclonal antibody. The clinical application of cellular immunoadsorption using the avidin-biotin system may prove useful in bone marrow transplantation.
生物素化抗体和抗生物素蛋白-琼脂糖6MB柱被用于一种从人骨髓中去除特定细胞群体的新方法。在一个模型系统中，将荧光素标记的Daudi淋巴母细胞与骨髓单个核细胞混合，并用倒置荧光显微镜对Daudi细胞的去除情况进行定量评估。使用与Bp32抗原（在Daudi细胞上表达）反应的生物素化单克隆抗体2H7进行处理，然后通过抗生物素蛋白-琼脂糖柱，可从骨髓中去除超过两个对数的Daudi细胞。通过先用未生物素化的单克隆抗体，然后用生物素化的山羊抗小鼠免疫球蛋白依次处理细胞，再通过抗生物素蛋白-琼脂糖柱，测试了另一种方法。通过造血祖细胞的定量回收，可从骨髓中消除多达三个对数的Daudi细胞。使用生物素化的山羊抗小鼠免疫球蛋白无需制备每种单克隆抗体的生物素缀合物。使用抗生物素蛋白-生物素系统进行细胞免疫吸附的临床应用可能在骨髓移植中证明是有用的。
