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 37^oC then shifted to 32^oC on day 05.

After 14 days, cell density reaches over 25 million per cell per milliliter, and Trastuzumab concentration collects around 04g/L.