(466c) Anti-Adhesive Glycosylated Proteins As a Technology for Improved Suspension Cell Culture in Bioreactors

Anti-Adhesive
Glycosylated Proteins as a Technology for Improved Suspension Cell Culture in
Bioreactors

            Mucins are brushlike
biopolymers found in biointerfaces, including joints
and on the surface of secretory glands (Figure 1A). Mucin biopolymers have
anti-adhesive properties that mediate lubrication, resist cell adhesion, and
protect cell surfaces, amongst other functions. Although mucin glycoproteins
have broad potential in medicine and biomaterials applications, their use has
been limited due to the challenge of biomanufacturing
sufficiently large quantities.  For
example, prokaryotic host production, despite its efficiency, cannot properly
reproduce the defining O-linked glycosylation of mucins. To address these
issues, we have created a protocol to efficiently produce and purify
structurally correct recombinant mucins from eukaryotic hosts, allowing us to
better understand the molecular basis of mucin structure and investigate
downstream applications, including their use as (1) biocompatible lubricants
and (2) anti-clumping agents in eukaryotic-cell bioreactors. In another
application, we engineered a membrane-anchored mucin, and observed that cells
expressing the engineered mucins spontaneously transition from adherent to
floating in the media, indicating decreased cell-cell and cell-matrix
interactions (Figure 1C, D). Remarkably, these cells displayed an increased
ability to survive in suspension compared to non-mucin expressing control cells
(Figure 1E). These results suggest that cell surface engineering with
genetically encoded mucins can improve the viability of eukaryotic cells,
possibly streamlining suspension adaptation and bioreactor performance.
Furthermore, the decreased cell-cell adhesivity of
engineered cells should minimize aggregation, a common problem that has limited
the use of human cell hosts in biomanufacturing.
Overall, our engineered cells have potential to become a new production
technology for biomanufacturing of complex biologics
and recombinant proteins with human-specific glycosylation.

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