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Updated: Nov 6, 2025

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
Published on: September 21, 2011
A simple and highly efficient counter-current chromatography method for the isolation of concentrated fractions of
1Kurnakov Institute of General & Inorganic Chemistry, Russian Academy of Sciences, Leninsky Prospekt 31, Moscow 119991, Russia.
Abstract:
A new modification of the conventional multiple sample loading (MSL) mode - sequential sample loading (SSL) - is suggested to enhance further the performance of the counter-current chromatography (CCC) separation processes. The sequential sample loading technique is simple and easy to implement: the continuous sample solution supply to a CCC column is alternated (interrupted) with short periods of the "pure" mobile phase supply. Periodic (batch) and continuous SSL CCC separations can be designed and implemented. In continuous processes, the sample solution loading is carried out in the form of separate series, consisting of a number of sequential sample solution loads. In this work, the modeling of the conventional multiple sample loading and the sequential sample loading counter-current chromatography is used to compare the two operating modes considered. Equations for the calculation of band profiles, the recovery yield and the purity are given. Equations are also derived permitting the calculation of the optimum operating parameters of the separation processes. It is shown that the use of sequential sample loading makes it possible to produce fractions of purified compounds with a much higher concentration than in the original sample solution. The simulations of the conventional multiple sample loading and the sequential sample loading counter-current chromatography separations are presented in "Mathcad" software.
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