LFIA: A Microfluidic Protein Interaction Analysis Method

LFIA：一种微流控蛋白质相互作用分析方法

• 批准号: 6935653
• 负责人: Rajan Kumar
• 金额: $ 24.76万
• 依托单位: GENOME DATA SYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-15 至 2007-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6935653
• 关键词: CD2 molecule; binding proteins; bioassay; bioengineering /biomedical engineering; biomedical automation; biomedical equipment development; biotin; chemical kinetics; evaluation /testing; high throughput technology; leukocyte adhesion molecules; microfluidics; protein binding; protein protein interaction; shear stress; technology /technique development; tumor necrosis factor alpha; video microscopy; viscosity

DESCRIPTION (provided by applicant): The primary goals of this SBIR project are 1) to develop a method to detect specific protein interactions by application of laminar flow, and 2) to develop a method to determine rate of protein interactions. Understanding of biomolecular interactions, especially protein-protein intractions, may hold the key to understand and treat complex diseases such as cancer, diabetes and neuro-degenerative and cardiovascular diseases. Novel methods for rapid identification of protein interactions and kinetic rate constant determination are needed. LFIA (laminar flow interaction analysis) will enable study of protein interactions in an innovative microfluidics assay. The Phase I application has the following three specific aims: 1. Investigate the feasibility of studying protein interactions using LFIA. We will fabricate and assemble microfluidics devices and associated instrumentation to perform LFIA and record experimental observations. 2. Characterize and optimize the performance of LFIA for protein interaction analysis. The specificity of protein interactions observed with LFIA will be confirmed. The compositions of the reagents and other LFIA components will be optimized to improve statistical significance of the results. 3. Design an instrument for automated LFIA. LFIA protocols will be optimized to enable automation of the assay. Preliminary specifications of an instrument to be fabricated in the Phase II project will be prepared. Although the application has specific emphasis on protein interactions, the method is generally applicable to any interaction between two analytes, including DNA, RNA, small molecules and carbohydrates. Phase II project will focus on increasing the robustness of LFIA process and development: of an automated instrument. Upon successful development, GDS will commercialize LFIA instrumentation and reagents, provide LFIA assays as a contract service, and develop biomolecular databases for licensing and for developing predictive models of human health and disease.

描述（由申请人提供）：该SBIR项目的主要目标是1）开发一种通过层流的应用来检测特定蛋白质相互作用的方法，以及2）开发一种确定蛋白质相互作用速率的方法。了解生物分子相互作用，尤其是蛋白质 - 蛋白质的作用，可能是理解和治疗复杂疾病的关键，例如癌症，糖尿病和神经性疾病和心血管疾病。需要快速鉴定蛋白质相互作用和动力学速率恒定测定的新方法。 LFIA（层流相互作用分析）将使在创新的微流体分析中研究蛋白质相互作用。 I阶段应用程序具有以下三个具体目标： 1。研究使用LFIA研究蛋白质相互作用的可行性。我们将制造和组装微流体设备和相关的仪器，以执行LFIA并记录实验观测。 2。表征和优化LFIA用于蛋白质相互作用分析的性能。将确认与LFIA观察到的蛋白质相互作用的特异性。试剂和其他LFIA成分的组成将被优化，以提高结果的统计意义。 3。设计自动LFIA的乐器。 LFIA协议将被优化以实现该测定的自动化。将准备在II期项目中制造的仪器的初步规格。尽管该应用特别强调蛋白质相互作用，但该方法通常适用于两个分析物（包括DNA，RNA，小分子和碳水化合物）之间的任何相互作用。 第二阶段项目将着重于提高LFIA过程和开发的鲁棒性：自动化仪器的鲁棒性。成功开发后，GDS将商业化LFIA仪器和试剂，提供LFIA分析作为合同服务，并开发用于许可和开发人类健康和疾病预测模型的生物分子数据库。