Development of Biosensor Technology for Protein Interactions

蛋白质相互作用生物传感器技术的发展

• 批准号: 8340622
• 负责人: PETER SCHUCK
• 金额: $ 6.38万
• 依托单位: NATIONAL INSTITUTE OF BIOMEDICAL IMAGING AND BIOENGINEERING
• 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/8340622
• 关键词: Accounting; Affinity; Binding; Binding Sites; Biosensor; Carbohydrates; Chemicals; Collaborations; Computer software; Data; Development; Goals; Heterogeneity; Kinetics; Laboratories; Lectin; Length; Membrane Glycoproteins; Membrane Proteins; Methods; Microscopic; Modeling; Polymers; Process; Property; Proteins; Role; Site; Surface; Technology; Testing; Update; chemical property; macromolecule; new technology; sensor

In the last year, we have applied our method of computationally accounting for surface site heterogeneity to a study of a model protein immobilized with different chemical strategies and on different sensor surfaces. We found that the most uniform surface site distributions were achieved with linker layers of moderate length. We identified experimental conditions that impose increased mass transport limitations. As an additional result of these studies, we have released an updated version of our distributable SPR data anlaysis software. This now also includes a discrete conformational change model, which allows testing of how well the hypothesis of slow conformational changes after an initial binding step fits experimental multi-phasic surface binding data, in comparison with the alternative explanation of surface site distributions with polydisperse affinity and kinetic constants. This was applied to data from the laboratory of Dr. Birgit Helm. Finally, we have continued the collaboration with the laboratory of Dr. Michael Tarlov at NIST on the recognition of different carbohydrate moieties on glycoproteins by surface-immobilized lectins.

去年，我们应用了计算方法，该方法考虑了表面位点异质性的研究，以研究与不同化学策略固定的模型蛋白质和不同的传感器表面。 我们发现，使用中等长度的接头层实现了最均匀的表面位点分布。 我们确定了实验条件，这些条件施加了增加的质量运输限制。 作为这些研究的另一个结果，我们发布了可分发的SPR数据Anlaysis软件的更新版本。 现在，这还包括一个离散的构象变化模型，该模型允许测试初始结合步骤后缓慢构象变化的假设拟合实验性多重表面结合数据，而与polydisperse亲和力和Kinetic常数的替代说明相比。 这应用于Birgit Helm博士实验室的数据。 最后，我们继续与NIST的迈克尔·塔洛夫（Michael Tarlov）博士的实验室合作，以表面上粘液化的凝集素在糖蛋白上识别不同的碳水化合物部分。