SBIR Phase II: An Automated Platform for Rapid Discovery in Cell Biology

小型企业创新研究(SBIR)第二阶段项目的更广泛影响/商业潜力是开发一种快速、高效和可扩展的细胞工程技术，通过与液体处理机器人的集成可以轻松实现自动化。目前，细胞工程尤其是细胞工程在发现新的治疗药物方面存在瓶颈。将基因或其他材料输送到细胞的领域没有跟上基因改造和高通量筛选技术的进步。建议的平台将提供一种替代耗时和劳动密集型的转基因方法，包括慢病毒转导和基于试管的电穿孔，这些方法很难自动化。细胞工程技术的应用范围从细胞生理学的基础研究到细胞治疗的新靶点的发现。该平台将使科学家和临床医生能够更快速、更可靠地设计免疫和其他细胞，以发现新的治疗靶点和疗法。这个SBIR第二阶段项目的智力优势将是开发一个可扩展的、自动化的非病毒细胞工程平台，其运行速度可能比使用高通量液体处理的传统电穿孔快10,000倍。利用第一阶段开发的核心细胞工程技术，目标是开发一种在液体处理机器人上进行基因导入的自动化协议，与96或384孔板技术兼容。第一个目标是证明用于高通量细胞工程的细胞工程设备的可制造性。这方面的初步工作表明，这些设备可以注塑成型，从而在降低成本的同时提高了规模化生产的潜力。在第二阶段项目中，将开发细胞工程设备的注塑原型，以证明可制造性并确定大规模制造的成本(每年数百万个部件)。其次，有几个补充系统必须与液体处理设备集成，才能实现拟议的高通量电池工程。补充系统包括与96或384井阵列的每个样品相互作用的电源和配电歧管。在本项目中，这些系统将与细胞工程设备和自动化液体处理机器人集成在一起。第三，集成系统将用于生成一个大型的人类初级T细胞变异库，作为概念验证，以展示高通量细胞工程用于治疗靶点发现的潜力。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase II project is to develop a fast, efficient, and scalable cell engineering technology that is easily automated through integration with liquid handling robots. Currently, there is a bottleneck in the process of cell engineering, especially in the engineering of cells for discovery of new therapeutics. The field of delivery of genetic or other material to cells has not kept pace with advancements in genetic modification and high-throughput screening technologies. The proposed platform will offer an alternative to the time-consuming and labor-intensive methods of transfection including lentiviral transduction and cuvette-based electroporation, which are difficult to automate. Applications of cell engineering technology range from fundamental research in cell physiology to the discovery of new targets for cellular therapies. The platform will allow scientists and clinicians to more rapidly and reliably engineer immune and other cells for discovery of new therapeutic targets and therapeutics.The intellectual merit of this SBIR Phase II project will be to develop a scalable, automated, non-viral cell engineering platform with the potential to operate up to 10,000 times faster than conventional electroporation using high-throughput liquid handling. Using the core cell engineering technology developed in Phase I, the goal is to develop an automated protocol for gene transfection on a liquid handling robot compatible with 96 or 384 well plate technology. The first objective is to demonstrate the manufacturability of cell engineering devices for high-throughput cell engineering. Preliminary work in this area has shown that these devices can be injection molded, thus reducing cost while increasing the potential for production at scale. In the Phase II project, injection molded prototypes of the cell engineering devices will be developed to prove manufacturability and determine the cost to manufacture at scale (millions of parts per year). Second, there are several supplemental systems that must be integrated with a liquid handling apparatus to enable the proposed high-throughput cell engineering. Supplemental systems include a power source and power distribution manifold that interacts with each sample of the 96 or 384 well array. In this project, these systems will be integrated with the cell engineering devices and automated liquid handling robot. Third, the integrated system will be used to generate a large library of primary human T cell variants as proof-of-concept to demonstrate the potential for high-throughput cell engineering for therapeutic target discovery.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.