Evaluation of multi-well based system for antibody production using Chinese hamster ovary cell

Abstract

The increased use of antibodies and antibody fragments for human therapy is driving the search for rational approaches to accelerate bioprocess development. However, screening of the operating conditions of process parameters in large scale bioreactors for biopharmaceuticals industry is not cost effective. Using microwell plate as the 'microreactor' for screening process wrould be a solution to this. Hence, this study was carried out to evaluate multi-well based system by determining the growth rate and antibody expression of the Chinese hamster ovary (CHO) cells. In this study, two types of CHO cells have been seeded with different concentrations which were 7^10^ cell/ml and lxlO6 cell/ml. Wild type CHO cell and CHO DG44 were used in this study. The wild type CHO cell was used as a reference cell as it is a stable cell. The cell number wras calculated by using trypan blue exclusion method while glucose concentration was determined by 3. 5- Dinitrosalicyclic acid (DNS) assay. On the other hand, antibody production was determined by using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. For the cell growth rate and doubling time, CHO DG44 (lxlO6 cell/ml) has recorded the highest growth rate and shortest doubling time with (0.3301 ± 0.1536)/h and (2.61 ±1.66)h. Besides that the highest glucose concentration recorded by wild type CHO cell (lxlO6 cell/ml) and CHO DG44 (7*10^ cell/ml) was on day 0 of incubation with (1,817±0.1187)mg/ml and (1.819±0.9331)mg/ml. Meanwhile, CHO DG44 (1 * 106 cell/ml) has recorded the highest glucose concentration on day 1 with (1.61 l±0.1273)mg/ml. Lastly, the antibodyexpress ion by CHO DG44 was verified. Both batches of CHO DG44 have successfully expressed human IgG on day 2 until day 10. In conclusion, the CHO cell is possible to be cultured in the multi-well plate.