title: Monensin, a small molecule ionophore, can be used to increase high mannose levels on monoclonal antibodies generated by Chinese hamster ovary production cell-lines.
abstract: Asparagine-linked glycosylation of the constant region of monoclonal antibodies (mAbs) plays an important role in their stability and efficacy and is a critical product quality attribute that needs to be consistent between various process changes and production lots. Exact product quality match is also of the utmost importance for the development of biosimilar protein therapeutics. This poses a process development challenge since mAb glycosylation profiles can fluctuate easily with changes in process parameters. Therefore, there is a need to identify methods to modulate glycosylation levels on therapeutic antibodies during a production run in order to maintain consistent product quality profiles between different drug lots. Here, we demonstrate the use of a small molecule ionophore, monensin, to increase high mannose levels on multiple therapeutic human immunoglobulins (IgGs) in both plate-based small scale production models as well as in production bioreactors. This method is simple to implement and readily applicable for multiple production cell lines. Moreover, high mannose levels can be increased without significant negative impact on titer or cell culture performance. As such, monensin gives us a manipulable product quality lever.
authors.affiliation: Cell Sciences and Technology, Amgen, 1201 Amgen Court W, Seattle, Washington, 98119.
authors.forename: Sandhya
authors.initials: S
authors.lastname: Pande
authors.affiliation: Cell Sciences and Technology, Amgen, 1201 Amgen Court W, Seattle, Washington, 98119.
authors.forename: Ayu
authors.initials: A
authors.lastname: Rahardjo
authors.affiliation: Analytical Sciences, Amgen, 1201 Amgen Court W, Seattle, Washington, 98119.
authors.forename: Brittney
authors.initials: B
authors.lastname: Livingston
authors.affiliation: Cell Sciences and Technology, Amgen, 1201 Amgen Court W, Seattle, Washington, 98119. mmujacic@gmail.com.
authors.forename: Mirna
authors.initials: M
authors.lastname: Mujacic
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000378:metabolism*; D016466:CHO Cells / Q000187:drug effects* / Q000378:metabolism*; D003412:Cricetulus; D006031:Glycosylation; D006801:Humans; D007476:Ionophores / Q000378:metabolism*; D008358:Mannose / Q000378:metabolism*; D008985:Monensin / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D007476:Ionophores; D011994:Recombinant Proteins; D008985:Monensin; D008358:Mannose
keywords: Chinese hamster ovary cells; glycosylation; golgi; high mannose; monensin; monoclonal antibodies; product quality control