Engineering and Analysis of Pressure Sensitive Antibodies

这个项目正在研究使用静水压力作为一种温和有效的分离抗原-抗体复合物的手段，从而为在分离和/或传感应用中利用这种压力敏感性的策略设计提供合理的基础。拟开展的研究包括三个方面：(1)利用高压荧光光谱对一组抗体的压力诱导现象进行基本的液相热力学和动力学表征；(2)蛋白质工程研究，旨在了解介导压力效应的分子机制；(3)固定化抗体的热力学、动力学和压力循环研究，以制定大规模实际压力洗脱的操作策略。除了为压力敏感抗体的工程和利用策略的开发提供合理的框架外，本研究还可以促进对压力对蛋白质结构和功能影响的基本理解。＊＊＊

9503227 Yarmush This project is on research to study the use of hydrostatic pressure as a mild and effective means of dissociating antigen-antibody complexes, thus providing a rational basis for the design of strategies which utilize this pressure sensitivity in separation and/or sensing applications. The proposed studies include three aspects: (1) fundamental solution-phase thermodynamic and kinetic characterization of the pressure-induced phenomena for a panel of antibodies using high pressure fluorescence spectroscopy; (2) protein engineering studies directed at understanding the molecular mechanisms mediating pressure effects; and (3) thermodynamic, kinetic and pressure cycling studies with immobilized antibodies to develop operational strategies for practical pressure elution on a large scale. In addition to providing the rational framework for the development of strategies for engineering and utilizing pressure sensitive antibodies, this research could also result in advances in the fundamental understanding of pressure effects on protein structure and function. ***