Alvarez, N., Adam, G.C., Howe, J.A., Sharma, V., Zimmerman, M.D., Dolgov, E., Rasheed, R., Nizar, F., Sahay, K., Nelson, A.M. and Park, S. Viruses 16, no. 7 (2024): 1158.
- Center for Discovery and Innovation, Hackensack Meridian Health, 111 Ideation Way, Nutley, NJ 07110, USA
- Merck & Co., Inc., Rahway, NJ 07065, USA
The enzymatic activity of recombinantly expressed 3CLPro enzymes from different coronaviruses was measured using the following synthetic quenched FRET peptide: CP488-ESATLQSGLRKAK- (CPQ2)-NH2 (CPC Scientific, San Jose, CA, USA).
Yarovaya, Olga I., Aleksandr S. Filimonov, Dmitriy S. Baev, Sophia S. Borisevich, Anna V. Zaykovskaya, Varvara Yu Chirkova, Mariya K. Marenina et al. " Viruses 16, no. 2 (2024): 215.
Fluorescence was observed by an assay involving a synthetic fluorescently labeled peptide substrate of the type DabcylKTSAVLQ↓SGFRKME(Edans)NH2 (more than 95% purity, CPC Scientific Inc., Hangzhou, China) containing the site digested by the main protease of coronavirus. The fluorescent dye attached to the peptide substrate allows the FRET effect to be implemented [..]
Shurtleff, V.W., Layton, M.E., Parish, C.A., Perkins, J.J., Schreier, J.D., Wang, Y., Adam, G.C., Alvarez, N., Bahmanjah, S., Bahnck-Teets, C.M. and Boyce, C.W. Journal of Medicinal Chemistry 67, no. 5 (2024): 3935-3958.
- Center for Discovery and Innovation, Hackensack Meridian Health. 111 Ideation Way. Nutley, New Jersey 07110, United States.
- Merck & Co., Inc., Rahway, New Jersey 07065, United States.
The enzymatic activity of recombinantly expressed 3CLPro enzymes from different coronaviruses was measured using the following synthetic quenched FRET peptide: CP488-ESATLQSGLRKAK- (CPQ2)-NH2 (CPC Scientific, San Jose, CA.
Kikuchi, F., Ikeda, Z., Kakegawa, K., Nishikawa, Y., Sasaki, S., Fukuda, K., Takami, K., Banno, Y., Nishikawa, H., Taya, N. and Nakahata, T. Bioorganic & Medicinal Chemistry 93 (2023): 117462.
- Research, Takeda Pharmaceutical Company Limited, 26-1, Muraoka-Higashi 2-chome, Fujisawa, Kanagawa 251-8555, Japan
- Pharmaceutical Sciences, Takeda Pharmaceutical Company Ltd., 26-1, Muraoka-Higashi 2-chome, Fujisawa, Kanagawa 251-8555, Japan
5FAM-Abu-Gly-Asp-Asp-Asp-Lys-Ile-Val-Gly-Gly-Lys(CPQ2)-Lys-Lys-NH2 (purity: 97.2%, CPC Scientific, Inc.) was diluted with an assay buffer to prepare a 5.4 μM substrate solution.
Ikeda, Z., Kakegawa, K., Kikuchi, F., Itono, S., Oki, H., Yashiro, H., Hiyoshi, H., Tsuchimori, K., Hamagami, K., Watanabe, M. and Sasaki, M. Journal of Medicinal Chemistry 65, no. 12 (2022): 8456-8477.
- Research, Takeda Pharmaceutical Company Limited, 26-1, Muraokahigashi 2-chome, Fujisawa, Kanagawa 251-8555, Japan
Subsequently, 5FAM–Abu–Gly–Asp–Asp–Asp–Lys–Ile–Val–Gly–Gly–Lys(CPQ2)–Lys–Lys–NH2 (purity: 97.2%, CPC Scientific, Inc.) was diluted with an assay buffer to prepare a 2.1 μM substrate solution.
Bekdemir, Ahmet, Eden EL Tanner, Jesse Kirkpatrick, Ava P. Soleimany, Samir Mitragotri, and Sangeeta N. Bhatia Advanced Healthcare Materials (2022): 2102685.
Peptides [TDN-qFAM, C-(PEG)2-K(CPQ2)-GGGSRPfGG-5FAM] were obtained from Tufts University Peptide Core Facility or CPC Scientific, Inc (USA)
Mac, Quoc D., Anirudh Sivakumar, Hathaichanok Phuengkham, Congmin Xu, James R. Bowen, Fang-Yi Su, Samuel Z. Stentz et al. Nature Biomedical Engineering (2022): 1-15.
"FITC-labelled GzmB substrate peptides with internal quencher ((5FAM)aIEFDSG(K-CPQ2)kkc) were synthesized by CPC Scientific and used for all in vitro activity assays. [..] Quenched fluorescent peptides for screening candidate substrates ((5FAM)-{substrate}-(K-DABCYL)-amide"
Soleimany, Ava P., [..] Sangeeta N. Bhatia. Cancer Research 81, no. 1 (2021): 213-224.
All peptides were synthesized by CPC Scientific (Sunnyvale, CA) and reconstituted in dimethylformamide (DMF) unless otherwise specified. Sequences are provided in Supplementary Table S1.
Cazanave, S.C., Warren, A.D., Pacula, M., Touti, F., Zagorska, A., Gural, N., Huang, E.K., Sherman, S., Cheema, M., Ibarra, S. and Bates, J. Science Translational Medicine 13, no. 616 (2021): eabe8939.
All peptides were synthesized by CPC Scientific Inc. For fluorogenic assays, peptides were flanked with the fluorophore-quencher pair 5-FAM-CPQ2. For in vivo urine experiments, peptides were barcoded with Glu-Fib–derived and heavy isotope–labeled peptides. PEGylated peptides were pooled prepared before in vivo experiments and stored at 4°C in phosphate-buffered saline.
Zeming, K. K.; Lu, R.; Woo, K. L.; Sun, G.; Quek, K. Y.; Cheow, L. F.; Chen, C.-H.; Han, J.; Lim, S. L. Analytical Chemistry 2021.
The FRET substrates used in this project are: neutrophil elastase substrate ((Z-AAAA)2Rh110 Abs/Em:497/520 nm, Anaspec), Granzyme B substrate (5TAMRA-VGPDFGR-K(QSY7)-NH2 Abs/Em: 546/580, CPC scientific), and metalloproteinase substrate (QSY21-HGDQMAQKSK(Cy5)-NH2 Abs/Em:649/666 nm, CPC Scientific).
