Yoshinobu Konno

Enhancement of antibody production by the addition of Coenzyme-Q10

Recently, there has been a growing demand for therapeutic monoclonal antibodies (MAbs) on the global market. Because therapeutic MAbs are more expensive than low-molecular-weight drugs, there have been strong demands to lower their production costs. Therefore, efficient methods to minimize the cost of goods are currently active areas of research. We have screened several enhancers of specific MAb production rate (SPR) using a YB2/0 cell line and found that coenzyme-Q10 (CoQ10) is a promising enhancer candidate. CoQ10 is well known as a strong antioxidant in the respiratory chain and is used for healthcare and other applications. Because CoQ10 is negligibly water soluble, most studies are limited by low concentrations. We added CoQ10 to a culture medium as dispersed nanoparticles at several concentrations (Q-Media) and conducted a fed-batch culture. Although the Q-Media had no effect on cumulative viable cell density, it enhanced SPR by 29%. In addition, the Q-Media had no effect on the binding or cytotoxic activity of MAbs. Q-Media also enhanced SPR with CHO and NS0 cell lines by 30%. These observations suggest that CoQ10 serves as a powerful aid in the production of MAbs by enhancing SPR without changing the characteristics of cell growth, or adversely affecting the quality or biological activity of MAbs.

Advantage of Alagln as an Additive to Cell Culture Medium

Advantage of dipeptides, L-alanyl-L-glutamine (AlaGln) and L-alanyl-L-tyrosine (AlaTyr), for the use of cell culture was tested in this study. Addition of the dipeptide to cell culture medium attenuated viability decline of cells and enhanced production titer of monoclonal antibody (Mab). Differences in stability between amino acids and dipeptides may significantly impact on cell culture performance. For example, medium containing glutamine (Gln) produced more ammonia than that of AlaGln. Accumulation of ammonia resulted in reduction of Mab titer, showing the advantage of using AlaGln.

Controlling Fucosylation Levels of Antibodies with Osmolality During Cell Culture in Several Host Cell Lines

Since a cost of therapeutic Monoclonal antibodies (MAbs) is much higher than other compounds, it is critical to produce high-efficacy MAbs efficiently. One method is to increase the effectiveness of a MAb, which in turn affects antibody-dependent cellular cytotoxicity (ADCC) is related defucosylation level (deFuc%) of MAbs. Since deFuc% of MAbs must be regulated for their quality control, it leads to careful consideration of the type of host cell employed. Thus, it is quite important to grasp the effects of culture conditions on the deFuc% in each cell line for the launched on the market and development, except for like a Chinese hamster ovary cells (CHOs) with α-1,6-fucosyltransferase gene knock out (PotelligentTM, BioWa, USA). For the MAbs produced in a rat myeloma cells (YB2/0), we found that osmolality of the culture medium is the major determinant of the deFuc%. In addition, deFuc% was not affected by the type of osmolytes (NaCl, KCl, fucose, fructose, and mannitol). We succeeded in controlling the deFuc% of MAbs arbitrarily 45–85% by maintaining medium osmolality during cultures (perfusion and fed-batch). We found the same correlation between the deFuc% and the culture osmolality in NS0 and SP2/0 cells as the in the YB2/0 cells.

Controlling Fucosylation Levels of Antibodies with Osmolality during Cell Culture

Because defucosylation levels (deFuc%) of monoclonal antibodies (MAbs) must be regulated for quality control, it is quite important to analyze the factors that affect deFuc%. For the MAbs produced in the rat hybridoma cell line YB2/0, we found that osmolality of the culture medium is the major determinant of deFuc%. deFuc% was linearly correlated with osmolality, with r2 being as high as 0.92. In addition, deFuc% was not affected by the type of compound used for controlling osmolality (NaCl, KCl, fucose, fructose, creatine and mannitol). We succeeded in controlling deFuc% of MAbs by maintaining medium osmolality constant during cultures (for both perfusion and fed-batch cultures).