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    • 8. 发明授权
    • Monoclonal antibody methods and compositions specific for single
antigens in antigen aggregates
    • 单克隆抗体方法和抗原聚集体中单抗原特异性的组合物
    • US4675386A
    • 1987-06-23
    • US495728
    • 1983-05-19
    • Ivor RoystonJohn MajdaGayle Yamamoto
    • Ivor RoystonJohn MajdaGayle Yamamoto
    • C07K16/28A61K39/395C12N5/00C12N15/00C12P21/00
    • C07K16/28Y10S435/948Y10S530/808
    • Methods and compositions are provided for producing monoclonal antibodies capable of distinguishing between antigenic aggregates, which are differentiated by one or a small number of antigenic determinants. The method involves immunization of a mammal to produce spleen cells which are fused with a cell line which is stable in vitro. The hybridized cells are then dispensed into culture wells at a density level which encourages the production of one colony per well. Autologous antigenic aggregates are employed for screening to distinguish the antibodies which discriminate between the antigenic aggregates. Particularly, mice are hyperimmunized with a homogeneous population of T-cells, the immunized mice killed and the spleens removed. The spleen cells are fused with mouse myeloma cells and the resulting hybrids plated at a relatively low density in culture wells. Plates containing fewer than 30% hybrid positive wells are found to have primarily one colony per well. The cell supernatants are screened against autologous T- and B-cells and hybrid antibodies which do not bind to the B-cells are specific for T-cells.
    • 提供了用于产生能够区分由一个或少数抗原决定簇分化的抗原性聚集体的单克隆抗体的方法和组合物。 该方法包括免疫哺乳动物以产生与体外稳定的细胞系融合的脾细胞。 然后将杂交的细胞以鼓励每孔产生一个菌落的密度水平分配到培养孔中。 自体抗原聚集体用于筛选以鉴别鉴别抗原聚集体之间的抗体。 特别地,用同样的T细胞群体对小鼠进行超免疫,免疫的小鼠被杀死,脾脏被去除。 将脾细胞与小鼠骨髓瘤细胞融合,并将所得杂交体以相对较低的密度铺在培养孔中。 发现含有少于30%混合阳性井的板块每孔主要有一个菌落。 对自体T细胞和B细胞筛选细胞上清液,并且不结合B细胞的杂交抗体对T细胞是特异性的。