A Novel Adherence Technology for Ophthalmic Medications

DESCRIPTION: Most human therapeutic glycoproteins (biologics) are manufactured using mammalian cell lines that can add the human-type glycans required for their clinical efficacy. However, mammalian cells have some serious disadvantages - it takes a longtime to establish production lines, they can contain adventitious agents, and they can produce immunogenic glycans. The baculovirus-insect cell system (BICS) offers an attractive alternative because it is fast, safe, and already being used to manufacture two FDA-licensed vaccines. Despite these advantages, the BICS is not yet being used to manufacture non-vaccine biologics because it cannot produce the human-type N-glycans needed for their efficacy. Our previous research has shown that we can solve this problem by glycoengineering insect cells to humanize their N-glycosylation pathways. However, those academic efforts were not designed to create optimally glycoengineered insect cell lines capable of efficient human-type N-glycosylation at low cost. Thus, theoverall goal of this Lab to Marketplace project is to create new glycoengineered insect cell lines that can provide efficient human-type N-glycosylation without costly media supplements for use in commercial biologics manufacturing and basic biomedical research. We will achieve this goal by extending our previous efforts and glycoengineering insect cells in new and innovative ways under pristine, isolated, and well-documented conditions. More specifically, we will isolate three new insect cell lines designed to produce glycoproteins with distinct, human-type N-glycans commonly found on commercial biologics. We will produce and maintain these cell lines under pristine, isolated, and well-documented conditions. We will then produce cell banks, test them for contaminants, and extensively characterize the cell lines to facilitate future validation and commercialization efforts. We also will examine the feasibility of using the new glycoengineered insect cell lines as manufacturing platforms by comparing their production levels to the parental cell line using three different glycoprotein biologics with native N-glycan structures identical to those to be produced, by design, by one or more of the new insect cell lines. Finally, we will examine the human-type N-glycosylation efficiencies of our new insect cell lines by expressing and purifying each biologic and quantitatively profiling their N-glycan structures. GlycoBac, LLC has the exclusive license to key intellectual property protecting glycoengineered BICS. After creating the new cell lines described herein, demonstrating their capabilities, and, validating their safety and efficacy in Phase II, GlycoBac will sublicense these lines to multiple biotechnology companies and contract manufacturing organizations. GlycoBac also will provide open access to these new cell lines for the worldwide basic and applied biomedical research communities. Thus, the tools generated in this project will have a significant and broad impact on human medicine and diverse areas of basicbiomedical research requiring properly and/or predictably glycosylated recombinant glycoproteins. PUBLIC HEALTH RELEVANCE PUBLIC HEALTH RELEVANCE: Most recombinant protein drugs, or biologics, are manufactured using mammalian cells that can add the sugars required for these proteins to have their clinical effect. However, mammalian cell systems have some limitations and new biologics manufacturing platforms are needed to overcome this problem and to address the need for increased manufacturingcapacity. This project will extend an academic lab effort, which has demonstrated that genetically engineered insect cells are potentially excellent alternative biologics manufacturing platforms, to the marketplace. New engineering strategies will be usedto create insect cell lines optimized for commercial applications under conditions that will facilitate their future validation as new biologics manufacturing tools.