Process development for Trastuzumab production from Chinese Hamster Ovary (CHO) in laboratory scale
Abstract
Background: CHO cells have been commonly used in the production of recombinant protein. It is reported that around 70% of recombinant therapeutic proteins are produced in CHO cell systems and the market size exceeds 15£ billion per year. With the optimization of culture, conditions showed that CHO cell density could reach 18 million per cell per milliliter and a final product concentration of 01 - 05 g l-1. Therefore, the production process development on a laboratory scale plays an important role in identifying the optimal conditions suitable, which can be applied to produce Trastuzumab on a industrial scale. In this study, Trastuzumab-producing cell lines were investigated in step wise: culture medium, percentage of supplements, culture temperature and shaking speed. The result showed that the optimal conditions were as follows: cells were shaken at 100rpm in ActiPro medium supplemented with Cell Boost 7a/7b (ratio 1:0.1%) daily from day 03, the initial growth temperature was 37oC then shifted to 32oC on day 05.
After 14 days, cell density reaches over 25 million per cell per milliliter, and Trastuzumab concentration collects around 04g/L.