Continuous cell screening using iso-dielectric separation

使用等电介质分离进行连续细胞筛选

• 批准号: 7021022
• 负责人: Joel Voldman
• 金额: $ 14.13万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-30 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7021022
• 关键词: Escherichia coli; cell sorting; dielectric property; electric field; electrical conductance; high throughput technology; method development; microfluidics; particle; polystyrenes; technology /technique development

DESCRIPTION (provided by applicant): We propose to develop a new approach for separating cells for cell-based screens. Separation of cells based upon biological differences-known as cell screening-is fundamental to both basic biology and biotechnology. These screens are an instrumental part of developing therapeutics and other biomolecles, and as such strongly impact human health. For both basic biology and biotechnology, one is interested in perturbing cells-to study or improve biosynthetic pathways-and then screening for desirable mutants in the population. These desirable mutants may produce new biomolecules or produce existing biomolecules more efficiently. In order to screen, one must have an experimental technique that can discern the phenotype of interest. The overall ability to study or engineer these organisms using screens is thus fundamentally dependent on our ability to measure. For biotechnology in particular, screening cells for biomolecule production typically requires development of a new assay for each biomolecule, even though the separation metric (e.g., production) is the same. Instead, we propose a new generic approach for separating cells based upon biomolecule production that is high-throughput, continuous, and real-time. This approach exploits the differences in electrical properties that exist between cells producing different amounts of biomolecule. Our hypothesis is that accumulating significant amounts of biomolecules will decrease cytoplasmic conductivity and permittivity in a way that we can exploit for particle separation. Our approach involves creating proportional spatial gradients of electric- field intensity and liquid conductivity, which causes cells to separate in space where their electrical properties match those of the liquid. We call this approach iso-dielectric separation (IDS). Our goal for this R21 proposal is to validate and characterize this approach with both test particles and bacteria. The relevance of this project to public health is that we are creating technology to better evaluate cells that produce biomolecules, including therapeutics. This has the potential to both increase the class of biomolecules available as well as more efficiently find & produce them.

描述（由申请人提供）：我们建议开发一种新方法，以分离基于细胞的屏幕的细胞。基于生物学差异的细胞分离，称为细胞筛选 - 这是基本生物学和生物技术的基础。这些屏幕是开发治疗剂和其他生物分子的重要组成部分，因此会对人类健康产生强烈的影响。对于基本生物学和生物技术，人们对研究或改善生物合成途径的细胞感兴趣，然后筛选人群中理想的突变体。这些理想的突变体可能会产生新的生物分子或更有效地产生现有的生物分子。为了筛选，必须具有一种可以辨别感兴趣表型的实验技术。因此，使用筛选研究或设计这些生物的总体能力从根本上取决于我们的测量能力。 特别是对于生物技术，即使分离度量（例如生产）相同，筛选生物分子产生的细胞通常需要为每种生物分子开发新测定。取而代之的是，我们提出了一种基于高通量，连续和实时的生物分子产生分离细胞的新通用方法。这种方法利用产生不同量的生物分子的细胞之间存在的电性能的差异。我们的假设是，积累大量的生物分子将以我们可以利用颗粒分离的方式降低细胞质的传导率和介电常数。我们的方法涉及创建电场强度和液体电导率的比例空间梯度，这会导致细胞在其电性能与液体相匹配的空间中分离。我们称这种方法为ISO-DIELECTRIC分离（IDS）。我们对R21提案的目标是通过测试颗粒和细菌验证和表征这种方法。 该项目与公共卫生的相关性是，我们正在创建技术来更好地评估产生生物分子（包括疗法）的细胞。这既有可能增加可用的生物分子类别，又可以更有效地找到和生产它们。