This record provides bibliographic details and links to the original research. An editorial study breakdown is not available.
What the researchers found
NanoClick measures the cumulative cytosolic exposure of a peptide in a concentration-dependent manner. It combines two technologies: in-cell click chemistry (where a chemical tag on the peptide reacts with a partner inside the cell) and NanoBRET (a light signal that only fires when the peptide reaches the cytoplasm).
The assay was validated using known cell-penetrating peptides and correlated with actual cellular activity using a p53/MDM2 model system (a well-studied protein-protein interaction in cancer). This confirms NanoClick measures functionally relevant cell penetration.
With minimal changes to the peptide sequence, NanoClick can detect entry via different mechanisms: endocytosis, direct membrane translocation, or passive permeability. This versatility is important because different peptides enter cells by different routes.
Why it matters
Macrocyclic peptides can target protein-protein interactions that small molecules cannot reach, but they must get inside cells to work. The lack of a high-throughput permeability assay has been a critical bottleneck. NanoClick fills this gap and could accelerate peptide drug discovery significantly.
The numbers in context
Validated with known CPPs; correlated with p53/MDM2 system; measures endocytosis, translocation, passive permeability
How the study worked
Researchers developed the NanoClick assay using HeLa cells expressing NanoBRET components. They validated it with known cell-penetrating peptides and a p53/MDM2 inhibitor peptide system. The assay uses click chemistry-compatible tags added to test peptides with minimal sequence disruption. High-throughput screening capability was demonstrated.
Who was studied
HeLa cells and peptide panels (in vitro assay development)
What this study cannot tell us
The assay requires adding click chemistry tags to peptides, which could subtly alter their properties. It was demonstrated primarily with cyclic peptides; performance with other peptide types may vary. The assay measures cytosolic exposure but not subcellular localization. It was validated in one cell line (HeLa).
Read the original research
NanoClick: A High Throughput, Target-Agnostic Peptide Cell Permeability Assay.
ACS chemical biology, 16(2), 293-309
Citation
Peier, Andrea; Ge, Lan; Boyer, Nicolas; Frost, John; Duggal, Ruchia; Biswas, Kaustav; Edmondson, Scott; Hermes, Jeffrey D; Yan, Lin; Zimprich, Chad; Sadruddin, Ahmad; Kristal Kaan, Hung Yi; Chandramohan, Arun; Brown, Christopher J; Thean, Dawn; Lee, Xue Er; Yuen, Tsz Ying; Ferrer-Gago, Fernando J; Johannes, Charles W; Lane, David P; Sherborne, Brad; Corona, Cesear; Robers, Matthew B; Sawyer, Tomi K; Partridge, Anthony W. (2021). NanoClick: A High Throughput, Target-Agnostic Peptide Cell Permeability Assay.. ACS chemical biology, 16(2), 293-309. https://doi.org/10.1021/acschembio.0c00804