An efficient method to control high mannose and core fucose levels in glycosylated antibody production using deoxymannojirimycin

Deoxymannojirimycin (DMJ) reduces the complex type glycan level and increases the high mannose level, in a dose dependent manner, during the production of IgG1 in CHO cell lines. Using DMJ in IgG1 production process enables tight control of core fucosylat. The effect of DMJ on complex and high mannose levels of IgG1 expressed in CHO cell. Red square- High Mannose; Blue diamond- Complex. [Display omitted] •Deoxymannojirimycin (DMJ) was used to modulate the glycosylation pattern of IgG1.•The effect of DMJ was shown in two different CHO host cell lines producing IgG1.•DMJ can be used in IgG1 manufacturing to control core fucose level and effect ADCC.•DMJ might be affecting Galactosyltransferase activity. Glycosylation on the Fc region of recombinant Immunoglobulin G (IgG) therapeutic antibodies is a critical protein quality attribute which may affect the efficacy and safety of the molecule. During the development of biosimilar therapeutics, adjustment of the glycosylation profile is required in order to match the reference innovator profile. Deoxymannojirimycin (DMJ), a known inhibitor of mannosidase, was used in this study to modulate the glycosylation pattern of antibodies. The effect of DMJ, at concentrations of 5 μM - 500 μM, on non-fucosylated glycoform levels was tested in the biosynthesis processes of two different IgG1 (IgG1 #A and IgG1 #B) using two Chinese hamster ovary (CHO) cell lines (CHO-DXB-11 and CHOK1SV, respectively) in Erlenmeyer flasks and in lab scale bioreactors. DMJ affected glycan forms in a dose response manner. At the highest concentration tested, DMJ reduced N-linked complex glycoform and core fucose levels by 15 and 14 fold, respectively, and increased high mannose level by 21 fold. 10 μM DMJ decreased IgG1 #A core fucose level in CHO-DXB-11 from 92% to 73% and increased high mannose level from 4% to 22% in Erlenmeyer flasks. Furthermore, in lab scale bioreactors, 15 μM DMJ decreased IgG1 #A core fucose level from 95% to 84% and increased high mannose level from 3% to 13%. Core fucose level of IgG1 #B in CHOK1SV was decreased from 81% to 73% using 10 μM DMJ in lab scale bioreactors while high mannose was increased from 6% to 15%. While affecting core fucose and high mannose levels, DMJ decreased maximum viable cell concentration by 16% and did not significantly affect cell productivity (less than 10%). This study demonstrated that DMJ can enable the control of core fucosylated and high mannose levels of IgG1 antibodies in a defined