The Study On Innovation And Scale-up Of Recombinant Human Interleukin-2 Purification Process

Interleukin-2 is a kind of cytokine,which can regulate the proliferation and differentiation of B cells,urge the fast proliferation of T cells and the NK cells as well.Currently the Interleukin-2 product in the market is mainly Recombinant Human Interleukin-2 expressed by gene engineering bacteria.The manufacturing process starts from the gene engineering bacteria through fermentation combined large scale separation and purification technology,finally gets the concentrated Recombinant Human Interleuking-2 and then the finished product.To meet the market development,we carried out the study over Recombinant Human Interleukin-2 manufacturing status and capacity upgrade.The capacity upgrade can be done in two aspects,mainly by upgrading the facilities used in the bacteria breaking-up sector and amplifying the down stream purification scale in order to reach the target of capacity improvement.The details of the study describe as below:1.Cell breaking-up technique innovation and scale-upTo solve the problem of insufficient processing by current breaking-up technique,in this study we adopted high pressure homogenizer and defined superior process condition parameters:homogenization pressure at 55MPa,breaking-up twice,initial temperature not higher 14? and cooling during the whole homogenization process.Under the above experiment condition,the results showed the best in the aspects of cell breaking-up rate,purification and unit time processing amount.Compared with the current technology,the purity increased by 9.2%,temperature rise rate reduced by 63.7%and unit time processing amount improved 27 times.2.Chromatography purification scale-upIn this study we evaluated the influence of chromatography column specification over the purification productivity and finally concluded that by using the column of ?20cm×80cm with a linear flow rate at 0.19cm/min the target protein yield and purity kept the same while the unit time processing amount increased by 4 times compared with current technology.3.Renaturation process scale-upIn this study,we evaluated the influence of different renaturation scale and time over the renaturation efficiency in the direct dilution renaturation,and defined 10L renaturation volume and 5 hours renatuation time as the superior renaturation condition.Compared with the current technology the biological activity and unit time processing amount both had obvious increase with a stable purity(biological activity and specific activity both increased by 16.8%,and unit time processing amount increased by 3.7 times).