#C1210N 3CLPro L50F + E166A + L167F Expression Kit

3CL^pro (M^pro) L50F + E166A + L167F mutation Expression Kit

In biochemical analysis, nirmatrelvir’s IC₅₀ value was found to be 72-fold greater in the presence of the L50F, E166A, L167F mutation (Jochmans et al. 2023). In a HEK293T cell-based 3CL^pro fluorescent reporter assay nirmtrelvir EC₅₀ increased from .96uM to 27uM with this mutation (Jochmans et al. 2023). These mutations occur independently in patients and have emerged collectively when passing the virus under nirmatrelvir selection (Jochmans et al. 2023; Shu and McCauley 2017). Furthermore, a FRET-based enzymatic assay demonstrated a 51.9-fold increase in IC₅₀ values for nirmatrelvir in the presence of this mutation (Huang et al. 2023).

• Identify protease inhibitors that maintain efficacy against the L50F + E166A + L167F 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 + E166A + L167F in BacMam
• Sodium Butyrate, 500 mM in H₂O

References:

Huang, C., Shuai, H., Qiao, J., Hou, Y., Zeng, R., Xia, A., Xie, L., Fang, Z., Li, Y., Yoon, C., Huang, Q., Hu, B., You, J., Quan, B., Zhao, X., Guo, N., Zhang, S., Ma, R., Zhang, J., … Yang, S. (2023). A new generation mpro inhibitor with potent activity against SARS-COV-2 omicron variants. Signal Transduction and Targeted Therapy, 8(1). https://doi.org/10.1038/s41392-023-01392-w

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