To establish the method of cell separation with domestic immuomagnetic beads, three methods were investigated. Direct method, SPA method and Biotin-Avidin method were applied to separate cell strain Hut-78 and CD4 positive cells. Separation rate of strain Hut-78 was more than 90% in direct method. Detachment rate with papain was over 95%. Cell activity was well retained. SPA method and Biotin-Avidin methods were also effective, but the direct method was superior to the other two techniques. Before separated by the direct method, CD4 positive cells constituted 46.4% +/- 6.4% of mononuclear cells (MNC), but in eliminated suspension there was only 6.2% +/- 2.3% CD4 positive cells left. In the separated part, 80.6% +/- 7.2% of the cells combined with the beads. It is concluded that the direct method in separating cells had high sensitivity and specificity.