High-performance liquid chromatography of amino acids, peptides and proteins. CXXXII. Optimisation of operating parameters for protein purification with chromatographic columns.
In large-scale chromatography, process optimisation is one of the key elements for success. This paper presents a method for determining the optimum operating parameters for affinity and ion-exchange chromatographic columns when used for protein purification. Based on a mathematical model developed as part of our association investigations, computer programs have been developed to describe the dynamic relationships acting within the chromatographic system. Two basic operating parameters, the flow-rate and the effluent concentration at which the adsorption stage is terminated, can be optimised to give a maximum production rate. The sample loading volume and the processing time then can be determined. The effect of washing conditions on the production rate and the yield is also discussed. Examples are given for a specific system where the optimisation is based on the yield and the percentage utilisation of the column capacity. Contour plots are generated to aid the determination of the range of controlling parameters, and to guide further system design.
Mao
Q M
QM
Department of Biochemistry, Monash University, Clayton, Vic., Australia.
Prince
I G
IG
Hearn
M T
MT
D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D007700:Kinetics; D008024:Ligands; D008956:Models, Chemical; D009113:Muramidase / Q000737:chemistry; D011506:Proteins / Q000302:isolation & purification*; D012984:Software; D014227:Triazines
D008024:Ligands; D011506:Proteins; D014227:Triazines; C012842:Cibacron Blue F 3GA; D009113:Muramidase