STTR Phase I: Affinity Chromatographic Cell Sorting (AFFICS) System for Simultaneous Multiple Cell Separation

这项小型企业技术转让（STTR）项目的更广泛影响/商业潜力将是开发一种细胞分选系统，允许分离两种或两种以上的细胞类型，用于治疗癌症和其他疾病的细胞疗法。从复杂材料（如血液）中纯化特定细胞群是细胞治疗制造的关键组成部分。然而，由于大多数临床级细胞纯化方案依赖于磁性颗粒与细胞的结合，因此有希望的细胞疗法面临将残留和潜在毒性磁性颗粒移植到人类受试者身上的风险。目前消除这种风险所需的步骤增加了细胞疗法制造的复杂性、时间和成本，限制了有前景的新细胞疗法的大规模应用。该项目将开发一种新的细胞分离技术，该技术不含磁性颗粒，可以很容易地扩展到临床规模生产高纯度和高存活率的细胞治疗药物所需的水平。目的是执行多目标细胞分选，同时使细胞免于污染标签。该STTR一期项目建议开发一种新型亲和色谱细胞分选（AFFICS）系统，该系统基于专有的可溶解水凝胶微珠，可直接从异质细胞环境中高效地同时分离两种或两种以上的细胞类型，而无需对细胞进行磁性或荧光标记。为了实现这一目标，目标是：1)设计一种基于柱的亲和色谱系统，该系统具有功能化的可溶水凝胶微珠，用于从复杂的细胞悬浮液中分离基于单表面标记的细胞，从而获得纯度和活力为85%的细胞捕获和靶细胞群。2)基于两个或多个表面标记物，开发多柱串联连接，用于大规模的细胞顺序分离。3)优化快速释放机制，在保持细胞表型和活力的同时，高效分离两个或多个细胞亚群。该系统将提供临床规模的多靶点细胞分选，同时使细胞免于残留的磁性或荧光亲和标签。

The broader impact/commercial potential of this Small Business Technology Transfer (STTR) project will be the development of a cell sorting system to allow separation of two or more cell types to be used in cell therapies for treatment of cancer and other diseases. Purification of specific cell populations from complex materials such as blood is a critical component of cell therapy manufacturing. However, since most protocols for clinical-grade cell purification rely on binding of magnetic particles to cells, promising cell therapies face the risk of transplanting residual and potentially toxic magnetic particles to human subjects. The steps currently required to eliminate this risk add complexity, time and cost to cell therapy manufacturing, limiting the wide-scale usefulness of promising new cell therapies. This project will develop a novel cell separation technology, free of magnetic particles, that can be readily scaled to levels required to enable clinical-scale manufacturing of highly pure and highly viable cell-based therapeutics. The aim is to perform multi-target cell sorting while simultaneously leaving cells free from contaminating tags.This STTR Phase I project proposes to develop a novel affinity chromatographic cell sorting (AFFICS) system, based on proprietary dissolvable hydrogel microbeads, for efficient simultaneous separation of two and more cell types directly from heterogeneous cell environments without requiring magnetic or fluorescent tagging of cells. To achieve this, the objectives are to 1) Design a column-based affinity chromatographic system with functionalized dissolvable hydrogel microbeads for single surface marker-based cell isolation from complex cell suspensions, leading to cell capture and target cell populations with 85% purity and viability. 2) Develop tandem connection of multiple columns for sequential cell separation based on two or more surface markers on a large scale. 3) Optimize the rapid release mechanism required for efficient simultaneous separation of two and more cell subpopulations while maintaining cell phenotype and viability. The proposed system will provide multi-target cell sorting at clinical scale while simultaneously leaving cells free of residual magnetic or fluorescent affinity tags.