Evaluating the use of Apo-neocarzinostatin as a cell penetrating protein

Moody, Paul; Burlina, Fabienne; Martin, Stephen R.; Morgan, Rachel E.; Offer, John; Smith, Mark E.B.; Molloy, Justin E.; Caddick, Stephen
April 2013
PEDS: Protein Engineering, Design & Selection;Apr2013, Vol. 26 Issue 4, p277
Academic Journal
Protein–ligand complex neocarzinostatin (NCS) is a small, thermostable protein-ligand complex that is able to deliver its ligand cargo into live mammalian cells where it induces DNA damage. Apo-NCS is able to functionally display complementarity determining regions loops, and has been hypothesised to act as a cell-penetrating protein, which would make it an ideal scaffold for cell targeting, and subsequent intracellular delivery of small-molecule drugs. In order to evaluate apo-NCS as a cell penetrating protein, we have evaluated the efficiency of its internalisation into live HeLa cells using matrix-assisted laser-desorption ionization-time-of-flight mass spectrometry and fluorescence microscopy. Following incubation of cells with apo-NCS, we observed no evidence of internalisation.


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