Recombinant manufacturing of multispecies biolubricants

Colville, M. J.; Huang, L.-T.; Schmidt, S.; Chen, K. J.; Vishwanath, K.; Su, J.; Williams, R.; Bonassar, L.; Reesink, H. L.; Paszek, M. J.

doi:10.1101/2024.05.05.592580

bioRxiv · 10.1101/2024.05.05.592580

Recombinant manufacturing of multispecies biolubricants

Colville, M. J.·Huang, L.-T.·Schmidt, S.·Chen, K. J.·Vishwanath, K.·Su, J.·Williams, R.·Bonassar, L.·Reesink, H. L.·Paszek, M. J.

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Abstract

Lubricin, a lubricating glycoprotein abundant in synovial fluid, forms a low-friction brush polymer interface in tissues exposed to sliding motion including joints, tendon sheaths, and the surface of the eye. Despite its therapeutic potential in diseases such as osteoarthritis and dry eye disease, there are few sources available. Through rational design, we developed a series of recombinant lubricin analogs that utilize the species-specific tissue-binding domains at the N- and C-termini to increase biocompatibility while replacing the central mucin domain with an engineered variant that retains the lubricating properties of native lubricin. In this study, we demonstrate the tissue binding capacity of our engineered lubricin product and its retention in the joint space of rats. Next, we present a new bioprocess chain that utilizes a human-derived cell line to produce O-glycosylation consistent with that of native lubricin and a purification strategy that capitalizes on the positively charged, hydrophobic N- and C-terminal domains. The bioprocess chain is demonstrated at 10 L scale in industry-standard equipment utilizing commonly available ion exchange, hydrophobic interaction and size exclusion chromatography resins. Finally, we confirmed the purity and lubricating properties of the recombinant biolubricant. The biomolecular engineering and bioprocessing strategies presented here are an effective means of lubricin production and could have broad applications to the study of mucins in general.

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Colville, M. J., Huang, L.-T., Schmidt, S., Chen, K. J., Vishwanath, K., Su, J., Williams, R., Bonassar, L., Reesink, H. L., Paszek, M. J.. 2024-05-05. Recombinant manufacturing of multispecies biolubricants. https://doi.org/10.1101/2024.05.05.592580

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