Preparative Liquid Chromatography Equipment for Fundamental Studies on Protein Purifications

首席研究员（PI）建议开展基础研究，以表征离子交换和亲和色谱系统中蛋白质的内在动力学和吸附参数。PI还建议建立设计蛋白质大规模色谱的方法。将研究几种具有很好表征分子量、等电点、氨基酸序列和三维结构的模型蛋白。以前为等压操作开发的计算机模拟将扩展到梯度洗脱系统。该模型将考虑非线性等温线、尺寸排斥、传质阻力、缓慢吸附-解吸动力学以及其他随时间变化的吸附现象，如蛋白质聚集和构象变化。模型和模拟结果将通过离子交换和亲和色谱柱实验进行验证。经过验证后，通过选择适当的设计和操作参数，如缓冲液pH、缓冲液类型、盐浓度、进料粒度、粒度和流速，模拟可用于设计更高效、更具成本效益的工艺。

The Principal Investigator (PI) proposes to carry out fundamental studies to characterize intrinsic kinetics and adsorption parameters for proteins in ion exchange and affinity chromatography systems. The PI also proposes to establish the methodology for designing large-scale chromatography for proteins. Several model proteins which have well characterized molecular weights, isoelectric points, amino acid sequences, and three dimensional structures will be studied. Computer simulations which have been previously developed for isocratic operations will be extended to gradient elution systems. Nonlinear isotherms, size exclusion, mass transfer resistances, slow adsorption-desorption kinetics, and other time-dependent adsorption phenomena such as protein aggregation and conformational changes will be considered in the model. The model and simulations will be validated with column experiments on ion exchange and affinity chromatography. After validation, the simulations can be used for designing more efficient and cost-effective processes by choosing appropriate design and operating parameters, such as buffer pH, types of buffer, salt concentration, feed size, particle size, and flow rate.