SBIR Phase II: Biomolecular Cell Injection With Nanofountain Probe Systems

这项小型企业创新研究（SBIR）二期项目旨在提高基于纳米山探针电穿孔（NFP-E）的新型单细胞技术的吞吐量。近年来，生物技术研究的进展显示出细胞重编程和极其敏感的医学诊断的希望，但这项研究需要有效、精确和温和的细胞转染，而目前缺乏一种强大的工具。NFP-E技术能够通过单细胞电穿孔实现剂量控制的生物分子、蛋白质或药物进入细胞的运输，从而满足这一需求。通过对目标细胞施加低电位，细胞膜上产生小孔隙，从而使分子以一种比目前使用的任何其他转染方法更有效、侵入性更小的方式进入细胞。该项目有三个主要目标：建立各种细胞类型和转染试剂的协议，制造与NFP-E耦合的微孔板，以及开发软件算法来自动校准和电穿孔控制。这些发展将创造一个强大的系统，具有卓越的过程控制和细胞活力，适用于相对高通量的单细胞转染应用。该项目的更广泛的影响/商业潜力源于这种新工具将为研究人员和生物技术公司提供前所未有的能力，用于操纵和询问细胞过程，有利于基础生物学研究和个性化医学应用的发展。本项目将结合纳米制造、微流体、生物物理学和分子生物学等方面，加强对基本电穿孔机制和单细胞分析技术的科学和技术理解。NFP-E工具将允许科学家将原代细胞用于研究应用，在实验室研究和人类疾病病理生理学之间提供更有意义的联系。此外，这种新工具可以通过启用生物技术的新功能，并为单细胞研究提供强大的商业工具，从而使个性化医疗的前景变得可行和实用，从而阐明疾病机制，集中药物发现工作，以及个性化疾病诊断和治疗。

This Small Business Innovation Research (SBIR) Phase II project is aimed at increasing the throughput of a novel single-cell technology based on nanofountain probe electroporation (NFP-E). Progress in biotechnology research in recent years has shown promise for cell reprogramming and extremely sensitive medical diagnostics, yet this research requires effective, precise, and gentle transfection of cells - for which a robust tool is currently lacking. The NFP-E technology is capable of filling this need by enabling single-cell electroporation for dosage-controlled transport of biomolecules, proteins, or drugs into a cell. By applying a low electric potential to a target cell, small pores in the cell membrane are generated, which allows for delivery of molecules into cells in a way that is more efficient and less invasive than any other method of transfection used today. This proposed project has three primary goals: to establish protocols for various cell types and transfection agents, to fabricate a microwell plate that couples with the NFP-E, and to develop software algorithms to automate alignment and electroporation control. These developments will create a robust system with exceptional process control and cell viability that is suitable for relatively high throughput single-cell transfection applications.The broader impact/commercial potential of this project stems from the unprecedented capabilities that the novel tool will provide to researchers and biotechnology companies for manipulation and interrogation of cellular processes, benefiting fundamental biology research and the development of personalized medicine applications. This project will enhance the scientific and technological understanding of fundamental electroporation mechanisms and single cell analysis techniques by combining nanofabrication, microfluidics, biophysics, and molecular biology aspects. The NFP-E tool will allow scientists to use primary cells for research applications, providing a more meaningful link between laboratory research and human disease pathophysiology. In addition, this new tool could make the promise of personalized medicine feasible and practical by enabling new capabilities in biotechnology and providing a robust commercial instrument for single-cell studies toward discoveries that elucidate disease mechanisms, focus drug discovery efforts, and personalize disease diagnosis and therapies.