Optimization of anion exchange chromatographic purification of adalimumab and evaluation of its combination with cation exchange method to improve purity.

Biopharmaceuticals intended for human use must meet exceptionally high purity requirements to avoid potential side effects. In order to improve the efficiency of the purification process, new types of ion exchange chromatography columns are being introduced. However, optimal assay methods for these new columns need to be developed, and therefore it is essential to create analytical methods that can effectively utilize them. The aim of this work was to develop an efficient anion exchange chromatography method for the purification of adalimumab, for which a Tosoh Skillpak NH2-750 column was used. First, the pH and elution volume of the sample were optimized, which allowed the determination of the appropriate method for optimizing the salt gradient at three different pH values. The protein content of the samples collected during the purification was examined with different setup parameters by size exclusion chromatography, and the size distribution and the amount of contaminants were determined by dynamic light scattering measurements. A combined method was also used, where first cation exchange chromatography was followed by anion exchange, which allowed the amount of contaminants to be reduced even further. Based on the results, an optimal anion exchange chromatography method was successfully developed, which not only achieved the appropriate protein concentration, but also significantly reduced the level of contaminants.