Small Scale Monoclonal Antibody Purification Platform

Abstract: The aim of this project is to transfer and implement a monoclonal antibody purification process that is traditionally done on an ÄKTAxpress to another machine called the ÄKTA pure. This is due to the fact that the ÄKTAxpress is being discontinued by the manufacturer and a replace-ment machine is needed for the time being. The project will include developing some new meth-ods that might also highlight the industry’s need for more advanced methods when an eventual replacement machine will reach the market. The purification process consists of firstly affinity chromatography followed by size exclusion chromatography. The implementation of this purification process was done with the help of ex-ternal Python-based control system Orbit. The implementation went well and the chromatograms produced showed clearly the load, wash and elution stages of the respective columns. Three pumps were used to load and wash the seri-ally connected columns. Some new methods that were created using Orbit was concentration-based pooling and sample change depending on sample volume. While testing the system during longer runs, the process yields were constant and therefore the system was considered quite robust. Another proof of concept implementation of the purification process was developed that used parallel flow paths with the aim to shorten overall process time. Instead of using three pumps to load and wash all columns, two pumps were connected to the affinity column and one to the size exclusion column. Using a special kind of valve, these two flow paths were connected to allow loading the size exclusion column from the eluate of the affinity column. This did indeed in-crease the process productivity by roughly 30%. The ÄKTA pure has a larger sample loading capacity per machine than the ÄKTAxpress. In addition, it also has a larger fraction collector. With the possibility to run the process both serial-ly and in parallel, it is concluded that the ÄKTA pure is more than capable to replace the ÄKTAxpress.