Measurement of protein associations via static light scattering

通过静态光散射测量蛋白质关联

• 批准号: 7593447
• 负责人: Allen P Minton
• 金额: $ 16.84万
• 依托单位: NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7593447
• 关键词: Annual Reports; Computer software; Condition; Cooperative Research and Development Agreement; Data; Data Analyses; Equilibrium; Future; Hour; Individual; Laboratories; Measurement; Modeling; Production; Proteins; Pump; Syringes; System; Techniques; Technology; Testing; Time; Validation; Work; base; concept; data acquisition; design; detector; light scattering; nephelometry; novel; research study

We have been testing pre-production versions of a system designed by Wyatt Technology Incorporated to facilitate composition-gradient static light scattering measurements. The system, based upon a concept developed previously in our laboratory (see annual reports of the previous two years), consists of a pump module containing three syringe pumps and an inline degasser, and a software package that is used to control the syringes to create composition gradients, to collect data from light scattering and concentration detectors, and to analyze the data in the context of models for equilibrium self-association of a single protein and hetero-association of two proteins. The hardware and software were loaned to us by Wyatt Technology under terms of CRADA DK-05-0104. Preliminary results indicate that with this system one can characterize and quantify equilibrium self-associations of each of two individual protein species and hetero-associations of the two proteins in a single experiment requiring only about 2-3 mg of each protein and less than one hour of data acquisition time.

我们一直在测试 Wyatt Technology Incorporated 设计的系统的预生产版本，以促进成分梯度静态光散射测量。 该系统基于我们实验室之前开发的概念（参见前两年的年度报告），由一个包含三个注射泵和一个在线脱气器的泵模块以及一个用于控制注射器以产生成分梯度，从光散射和浓度检测器收集数据，并在单个蛋白质的平衡自缔合和两个蛋白质的异质缔合模型的背景下分析数据。 硬件和软件由 Wyatt Technology 根据 CRADA DK-05-0104 条款借给我们。 初步结果表明，利用该系统，人们可以在一次实验中表征和量化两个单独蛋白质种类中的每一个的平衡自缔合以及两个蛋白质的异质缔合，仅需要大约 2-3 毫克每种蛋白质和不到一小时的时间数据采集​​时间。