Prediction and Understanding of Protein Affinity

Cramer 0418413将蛋白质吸附的分子水平研究与宏观/过程规模相联系一直是生物分离领域的长期目标。该项目将专注于开发新的方法，以使在各种条件下的蛋白质结合亲和力的先验预测离子交换色谱系统。将采用高通量实验来积累大量色谱数据。将开发一种计算效率高的工具，将基本的水介导的相互作用与关键的表面位点，如带电配体的蛋白质表面相互作用的自由能的预测个别氨基酸和肽。新型分子描述符将被开发并用于先进的机器学习方法，以构建日益复杂的实验系统的模型。最初的实验和理论工作将集中在定义良好的表面，如自组装单分子层（SAMs）和多层膜。这些模型系统开发的技术和见解将被应用于更复杂的离子交换色谱系统。将进行实验以产生大量的线性和非线性色谱吸附数据，这些数据将构成两种互补色谱建模工作的基础。这些方法的协同作用提供了一个独特的机会，开发强大的预测工具和方法，以获得对蛋白质离子交换过程的基本物理学的重要见解。这将对设计基于高效离子交换色谱（IEC）的蛋白质分离过程的能力产生重大影响，并将适用于色谱以外的蛋白质吸附发挥重要作用的几个领域，如生物材料和药物输送。

Cramer0418413It has been a long standing goal in the bioseparations field to connect molecular-level investigations of protein adsorption to the macroscopic/process scale. This project will focus on the development of novel methods to enable the a priori prediction of protein binding affinity to ion-exchange chromatographic systems under a variety of conditions. High throughput experimentation will be employed to accumulate a large amount of chromatographic data. A computationally efficient tool will be developed that incorporates fundamental water-mediated interactions between individual amino acids and peptides with critical surface sites such as charged ligands for the prediction of protein-surface interaction free energies. Novel molecular descriptors will be developed and employed in advanced machine learning methods to construct models of increasingly complex experimental systems. Initial experimental and theoretical work will focus on well-defined surfaces such as self assembled monolayers (SAMs) and polyelectrolyte multilayers. The techniques and insights developed with these model systems will then be applied to more complicated ion-exchange chromatographic systems. Experiments will be carried out to produce a large set of linear and non-linear chromatographic adsorption data that will form the basis for two kinds of complementary chromatographic modeling work. The synergy of these methods provides a unique opportunity to develop powerful predictive tools and methods of gaining significant insight into the fundamental physics of the protein ion-exchange process. This will have a major impact on the ability to design efficient ion-exchange chromatography (IEC) based protein separation processes and will have applicability beyond chromatography to several fields where protein adsorption plays an important role such as biomaterials and drug delivery.