TOPIC 397 - FUNCTIONALLY CLOSED PURIFICATION AND ELUTION OF UNTOUCHED T CELLS USING CLEAVABLE MAGNETIC BEADS WITH RATCHETING CYTOMETRY

这项工作的目标是开发一种功能封闭、高通量的磁性细胞纯化系统，该系统可以实现：1)通过阳性选择直接从白细胞生物基质中纯化T细胞群；2)切割头细胞键，产生用于细胞治疗的未接触的T细胞。我们相信，一个功能封闭的系统，可以实现目标细胞群的高纯度阳性选择，将提高最终治疗细胞批次的效力和可重复性，并使治疗生产使用更细致和精致的细胞类型。在第一阶段，我们将使用Ferrologix现有的T细胞纯化盒和市售的可切割磁珠进行分离。白细胞/单血细胞分离标本将首先用抗cd3可切割磁珠标记，用Ferrologix棘轮细胞测定试剂盒分离，用头切割剂在试剂盒中孵育，T细胞从芯片上洗脱，而珠仍被捕获在试剂盒中。Ferrologix未触碰T细胞工作流程将与Miltenyi和StemCell Technologies的其他两个可切割头工作流程进行拆分样品方式的比较。工作流程将在处理时间、纯度、活力和磁珠成功切割的基础上进行比较。

The goal of this effort is to develop a functionally closed, high throughput magnetic cell purification system which can achieve 1) purification of T cell populations directly from a leukopak biomatrix via positive selection and 2) cleavage of bead-cell bonds resulting in untouched T cells for cell therapy production. We believe that a functionally closed system that can achieve high purity positive selection of target cell populations will improve potency and repeatability of the final therapeutic cell batches as well as enable therapeutic production using more nuanced and delicate cell types. In Phase I we will perform separations using Ferrologix existing T cell purification cartridge and commercially available cleavable magnetic beads. Leukopak/apheresis specimens will be first tagged with anti-CD3 cleavable magnetic beads, separated with Ferrologix ratcheting cytoemtry cartridge, incubated within the cartridge with bead cleaving agents, and T cell elution off chip while the beads remain trapped in the cartridge. Ferrologix untouched T cell workflow will be compared in split sample fashion with two other cleavable bead workflows from Miltenyi and StemCell Technologies. Workflows will be compared on the basis of processing time, purity, viability, and successful cleavage of magnetic beads.