The molecular weight and concentration of dextran sulfate affect cell growth and antibody production in CHO cell cultures.
To investigate the effect of dextran sulfate (DS), a widely used anti-aggregation agent, on cell growth and monoclonal antibody (mAb) production including the quality attributes, DS with the three different MWs (4,000 Da, 15,000 Da, and 40,000 Da) at various concentrations (up to 1 g/L) was added to suspension cultures of two different recombinant CHO (rCHO) cell lines producing mAb, SM-0.025 and CS13-1.00. For both cell lines, the addition of DS, regardless of the MW and concentration of DS used, improved cell growth and viability in the decline phase of growth. However, it increased mAb production only in the CS13-1.00 cells. Among the three different MWs, 40,000 Da DS was most effective in attenuating cell aggregation during the cultures of CS13-1.00 cells, and showed the highest maximum mAb concentration. For SM-0.025 cells, it significantly decreased specific mAb productivity, particularly at a high concentration of DS. Overall, DS addition did not negatively affect the quality attributes of mAbs (aggregation, charge variation, and glycosylation), though its efficacy on mAb quality depended on the MW and concentration of DS and cell lines. For both cell lines, the addition of DS did not affect N-glycosylation of mAbs and decreased basic charge variants in mAbs. For CS13-1.00 cells, the mAb monomer increased with the addition of 40,000 Da DS at 0.3-1.0 g/L. Taken together, to maximize the beneficial effect of DS addition on mAb production, the optimal MW and concentration of DS should be determined for each specific rCHO cell line. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1113-1122, 2016.
Department of Biological Sciences, KAIST, 373-1 Kusong-Dong, Yusong-Gu, Daejon, 305-701, Republic of Korea.
Jin
J
Hyoung Park
New Drug Development Center, Cheongju, Republic of Korea.
Myung
M
Sin Lim
New Drug Development Center, Cheongju, Republic of Korea.
Ju
J
Rang Woo
New Drug Development Center, Cheongju, Republic of Korea.
Jong
J
Won Kim
Department of Biological Sciences, KAIST, 373-1 Kusong-Dong, Yusong-Gu, Daejon, 305-701, Republic of Korea. gyunminlee@kaist.ac.kr.
Gyun
G
Min Lee
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000917:Antibody Formation; D016466:CHO Cells; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003412:Cricetulus; D016264:Dextran Sulfate / Q000737:chemistry / Q000494:pharmacology*; D008970:Molecular Weight
D000911:Antibodies, Monoclonal; D016264:Dextran Sulfate
CHO cell; N-glycosylation; charge variation; dextran sulfate; monoclonal antibody