Ophir, Y.; Wong, J.; Haddad, K. R.; Huuskonen, A.; Karmaker, A.; Gore, V.; Jung, S.; Oloumi, A.; Liu, Y.; Fu, J.; Zhang, L.; Huang, P.; Minami, S. A.; Garimella, S. S.; Thyagatur, A.; Zaini, P. A.; Vitikainen, M.; Tchelet, R.; Valbuena, N.; Fuerst, T.; Korkmaz, E.; Falo, L. D.; Balmert, S. C.; Mahendiratta, S.; Emalfarb, M.; Shah, P.; Siegel, J. B.; Dandekar, A. M.; Chen, X.; Lebrilla, C. B.; Faller, R.; Saloheimo, M.; McDonald, K. A.; Nandi, S.

The COVID-19 pandemic demonstrated a pressing need for rapid, adaptive, and scalable manufacturing of vaccines and reagents. With the transition into an endemic disease and rising threats of other emerging pandemics, production of these biologicals requires a stable and sustainable supply chain and accessible distribution methods. In this study, we demonstrate the strength of an engineered filamentous fungal platform, Thermothelomyces heterothallica C1, for high volumetric productivity of the full-length spike glycoprotein. Spike protein produced in this system is highly thermostable and immunization of mice with spike made in C1 or mammalian platforms resulted in a similar humoral response. Additionally, it was shown that the native N-glycan profile can be redecorated with complex sialylated structures, if necessary, resulting in a more human-like glycan profile, without impacting binding characteristics as shown experimentally and in simulations. Through extensive physicochemical analysis, the C1 produced spike performs similarly to spike proteins produced in other commercially available systems. The data presented is evidence that C1 can be a strong platform for production of complex glycosylated recombinant proteins such as subunit antigen vaccines.