#C1201N 3CLPro L50F Expression Kit

3CL^pro (M^pro) L50F mutation Expression Kit

L50F is a compensatory substitution that aids in maintaining the virus’s fitness (Jochmans et al. 2023; Zhou et al. 2022; Iketani et al. 2023). In a recombinant SARS-CoV-2 assay conducted in huh7-ACE2 cells, L50F has a 4.2-fold resistance to nirmatrelvir (Iketani et al. 2023). Structurally, the L50F mutation is located in the S2 subsite of 3CLpro and demonstrates increased catalytic activity compared to wild-type 3CLpro in kinetic assays (Duan et al. 2023). This mutation has been detected in patients and documented in the GISAID database (Shu and McCauley 2017).

• Identify protease inhibitors that maintain efficacy against the L50F mutation
• Easy co-expression with our 3CLglow and 3CLglowUp sensors
• Cost effective screening with minimal safety requirements (BSL-1)

Materials in the Kit:

• 3CL^pro L50F in BacMam

• Sodium Butyrate, 500 mM in H₂O

References:

Duan, Y., Zhou, H., Liu, X., Iketani, S., Lin, M., Zhang, X., Bian, Q., Wang, H., Sun, H., Hong, S. J., Culbertson, B., Mohri, H., Luck, M. I., Zhu, Y., Liu, X., Lu, Y., Yang, X., Yang, K., Sabo, Y., … Yang, H. (2023). Molecular mechanisms of SARS-COV-2 resistance to Nirmatrelvir. Nature, 622(7982), 376–382. https://doi.org/10.1038/s41586-023-06609-0

Iketani, S., Mohri, H., Culbertson, B., Hong, S. J., Duan, Y., Luck, M. I., Annavajhala, M. K., Guo, Y., Sheng, Z., Uhlemann, A.-C., Goff, S. P., Sabo, Y., Yang, H., Chavez, A., & Ho, D. D. (2023). Multiple pathways for SARS-COV-2 resistance to Nirmatrelvir. Nature, 613(7944), 558–564. https://doi.org/10.1038/s41586-022-05514-2

Jochmans, D., Liu, C., Donckers, K., Stoycheva, A., Boland, S., Stevens, S. K., De Vita, C., Vanmechelen, B., Maes, P., Trüeb, B., Ebert, N., Thiel, V., De Jonghe, S., Vangeel, L., Bardiot, D., Jekle, A., Blatt, L. M., Beigelman, L., Symons, J. A., … Vandyck, K. (2023). The substitutions L50F, E166A, and L167F in SARS-COV-2 3clpro are selected by a protease inhibitor in vitro and confer resistance to Nirmatrelvir. mBio, 14(1). https://doi.org/10.1128/mbio.02815-22

Shu, Y., & McCauley, J. (2017). GISAID: Global initiative on sharing all influenza data – from vision to reality. Eurosurveillance, 22(13). https://doi.org/10.2807/1560-7917.es.2017.22.13.30494

Zhou, Y., Gammeltoft, K. A., Ryberg, L. A., Pham, L. V., Tjørnelund, H. D., Binderup, A., Duarte Hernandez, C. R., Fernandez-Antunez, C., Offersgaard, A., Fahnøe, U., Peters, G. H., Ramirez, S., Bukh, J., & Gottwein, J. M. (2022). Nirmatrelvir-resistant SARS-COV-2 variants with high fitness in an infectious cell culture system. Science Advances, 8(51). https://doi.org/10.1126/sciadv.add7197