TY - JOUR
T1 - Expression and characterization of SARS-CoV-2 spike protein in Thermothelomyces heterothallica C1
AU - Ophir, Yakir
AU - Wong, Justin H.
AU - Haddad, Katherine R.
AU - Huuskonen, Anne
AU - Karmaker, Anindya
AU - Gore, Varun
AU - Jung, Seongwon
AU - Oloumi, Armin
AU - Liu, Yiyun
AU - Fu, Jingxin
AU - Zhang, Libo
AU - Huang, Peishan
AU - Minami, Shiaki Arnett
AU - Garimella, Shruthi Satya
AU - Thyagatur, Anugraha
AU - Zaini, Paulo A.
AU - Vitikainen, Marika
AU - Tchelet, Ronen
AU - Valbuena, Noelia
AU - Fuerst, Thomas R.
AU - Korkmaz, Emrullah
AU - Falo, Louis D.
AU - Balmert, Stephen C.
AU - Mahendiratta, Saniya
AU - Emalfarb, Mark
AU - Shah, Priya S.
AU - Siegel, Justin
AU - Dandekar, Abhaya M.
AU - Chen, Xi
AU - Lebrilla, Carlito
AU - Faller, Roland
AU - Saloheimo, Markku
AU - McDonald, Karen A.
AU - Nandi, Somen
PY - 2025/10/24
Y1 - 2025/10/24
N2 - 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.
AB - 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.
KW - Bioproduction
KW - COVID-19
KW - SARS-CoV2
KW - Spike protein
KW - Thermothelomyces heterothallica C1
KW - Vaccines
UR - https://www.scopus.com/pages/publications/105017315927
U2 - 10.1016/j.vaccine.2025.127784
DO - 10.1016/j.vaccine.2025.127784
M3 - Article
AN - SCOPUS:105017315927
SN - 0264-410X
VL - 65
JO - Vaccine
JF - Vaccine
M1 - 127784
ER -
