title: Effect of chemical chaperone addition on production and aggregation of recombinant flag-tagged COMP-angiopoietin 1 in Chinese hamster ovary cells.
abstract: To investigate the effect of chemical chaperones on the production and aggregation of flag-tagged cartilage oligomeric matrix protein-Angiopoietin1 (FCA1) in recombinant Chinese hamster ovary (CHO) cells, CHO cells were cultivated in serum-free media with various chemical chaperones, 1 mM 4-phenylbutyrate (4-PBA), 200 mM proline, 3% glycerol, 2% dimethyl sulfoxide (DMSO), and without chemical chaperone as control. The addition of chemical chaperones enhanced FCA1 production and specific FCA1 productivity, q(FCA)(1). For example, the q(FCA)(1) at 200 mM proline was fourfold higher than that at control. Unlike q(FCA)(1), the aggregation of FCA1 was strongly affected by which chemical chaperone was added. The addition of 2% DMSO and 200 mM proline significantly reduced the proportion of aggregates, but the addition of 1 mM 4-PBA and 3% glycerol was hardly effective. The proportions of aggregates were 29.5 and 55.6% at 2% DMSO and 200 mM proline, respectively, whereas it was 79.6% at control. The exact mechanism how chemical chaperones affected the aggregation of FCA1 was not investigated in this study, and therefore, more extensive works will be needed to clarify why different chemical chaperones behaved differently in reducing the aggregation of FCA1. Among chemical chaperones tested, DMSO was the most effective one in regard to enhancing the production and reducing the aggregation of FCA1 in CHO cells.
authors.affiliation: Department of Biological Sciences, KAIST, 373-1 Guseong-Dong, Yuseong-gu, Daejeon 305-701, Korea.
authors.forename: Su-Jeong
authors.initials: SJ
authors.lastname: Hwang
authors.forename: Choon-Ju
authors.initials: CJ
authors.lastname: Jeon
authors.forename: Sung Min
authors.initials: SM
authors.lastname: Cho
authors.forename: Gyun Min
authors.initials: GM
authors.lastname: Lee
authors.forename: Sung Kwan
authors.initials: SK
authors.lastname: Yoon
mesh_terms: D042683:Angiopoietin-1 / Q000096:biosynthesis* / Q000737:chemistry; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D015335:Molecular Probes; D055503:Protein Multimerization / Q000187:drug effects*; D011994:Recombinant Proteins; D012997:Solvents / Q000494:pharmacology*
chemical_list: D042683:Angiopoietin-1; D015335:Molecular Probes; D011994:Recombinant Proteins; D012997:Solvents; D004121:Dimethyl Sulfoxide