Preparative Liquid Chromatography Equipment for Fundamental Studies on Protein Purifications

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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.