Zborowski M, Moore L R, Reddy S, Chen G H, Sun L, Chalmers J J
Department of Biomedical Engineering, Cleveland Clinic Foundation, OH 44195-5254, USA.
ASAIO J. 1996 Sep-Oct;42(5):M666-71. doi: 10.1097/00002480-199609000-00071.
Cells of identical physical properties that differ in the expression of surface proteins can be sorted conveniently using immunospecific stains conjugated to fluorescent, or magnetic, labels. Immunomagnetic cell sorting using commercial batch sorters offers advantages of high sorting capacity, high viability of sorted fractions, and high depletion rates; its disadvantages are low enrichment rate and batch processing. The authors developed and tested a continuous, flow-through magnetic cell sorter for small volume, experimental cell enrichment. Freshly isolated human peripheral lymphocytes were labeled using an immunofluoromagnetic sandwich consisting of mouse anti human CD8 monoclonal antibody-fluorescein conjugate and rat anti mouse polyclonal antibody-colloidal iron-dextran conjugate. A total of 2-3 min lymphocytes were sorted per hour using a saturation magnetic field of 1.334 T and a five channel sorter. The fluorescent cells were distributed among the channels in relation to their fluorescence intensity and magnetic susceptibility. The purity (68-85%) and enrichment rates (16-34x) were comparable to those of commercial batch magnetic separators; sorting capacity and recovery of the enriched fractions (up to 32%) were limited by the small scale of the sorter. Future direction is focused on increasing the resolution, recovery, and sorting capacity of the enriched fractions, and testing the sorter on other cell systems.
表面蛋白表达不同但物理性质相同的细胞，可以使用与荧光或磁性标签偶联的免疫特异性染料方便地进行分选。使用商用批量分选仪进行免疫磁珠细胞分选具有分选能力高、分选组分活力高和去除率高的优点；其缺点是富集率低和分批处理。作者开发并测试了一种用于小体积实验性细胞富集的连续流通式磁细胞分选仪。使用由小鼠抗人CD8单克隆抗体 - 荧光素偶联物和大鼠抗小鼠多克隆抗体 - 胶体铁 - 葡聚糖偶联物组成的免疫荧光磁珠夹心对新鲜分离的人外周血淋巴细胞进行标记。使用1.334 T的饱和磁场和五通道分选仪，每小时可分选2 - 3分钟的淋巴细胞。荧光细胞根据其荧光强度和磁化率分布在各通道之间。纯度（68 - 85%）和富集率（16 - 34倍）与商用批量磁分离器相当；分选能力和富集组分的回收率（高达32%）受分选仪规模较小的限制。未来的方向集中在提高富集组分的分辨率、回收率和分选能力，以及在其他细胞系统上测试该分选仪。
