The influence of degree of labelling upon cellular internalisation of antibody-cell penetrating peptide conjugates.

Autor: Pringle TA; School of Natural and Environmental Sciences, Newcastle University Newcastle Upon Tyne UK james.knight2@newcastle.ac.uk., Coleman O; School of Natural and Environmental Sciences, Newcastle University Newcastle Upon Tyne UK james.knight2@newcastle.ac.uk., Kawamura A; School of Natural and Environmental Sciences, Newcastle University Newcastle Upon Tyne UK james.knight2@newcastle.ac.uk.; Chemistry Research Laboratory, Department of Chemistry, University of Oxford Oxford UK., Knight JC; School of Natural and Environmental Sciences, Newcastle University Newcastle Upon Tyne UK james.knight2@newcastle.ac.uk.; Newcastle University Centre for Cancer, Newcastle University Newcastle Upon Tyne UK.
Jazyk: angličtina
Zdroj: RSC advances [RSC Adv] 2022 Sep 28; Vol. 12 (43), pp. 27716-27722. Date of Electronic Publication: 2022 Sep 28 (Print Publication: 2022).
DOI: 10.1039/d2ra05274a
Abstrakt: Antibody-based agents are increasingly used as therapeutics and imaging agents, yet are generally restricted to cell surface targets due to inefficient cellular internalisation and endosomal entrapment. Enhanced cell membrane translocation of antibodies can be achieved by the covalent attachment of cell-penetrating peptides, including the HIV-1-derived transactivator of transcription (TAT) peptide. This study evaluated the cellular internalisation properties of five anti-HER2 Herceptin-TAT conjugates with degrees of TAT labelling (DOL TAT ) ranging from one to five. Herceptin-TAT conjugates were synthesised via a strain-promoted alkyne-azide cycloaddition reaction, characterised by UV-vis spectroscopy, MALDI-TOF, and gel electrophoresis, then radiolabelled with zirconium-89 to permit measurement of cellular internalisation by gamma counting. [ 89 Zr]Zr-DFO-Her-TAT (0-5) conjugates were isolated in high radiochemical purity (>99%) and exhibited high stability in murine and human serum over 7 days at 37 °C. Significant increases in cellular internalisation were observed for [ 89 Zr]Zr-DFO-Her-TAT conjugates with DOL TAT values of 2 and above in SKBR3 (high HER2) cells over 48 h, in contrast to low-level non-specific uptake in MDA-MB-468 (low HER2) cells that did not increase over time. Notably, [ 89 Zr]Zr-DFO-Her-TAT conjugates with DOL TAT of 3, 4, and 5 reached uptake values in SKBR3 cells of 5, 6, and 8% of the applied dose at 48 h respectively, representing 9, 10, 14-fold increases relative to the TAT-free control conjugate, [ 89 Zr]Zr-DFO-Her-TAT (0) .
Competing Interests: There are no conflicts to declare.
(This journal is © The Royal Society of Chemistry.)
Databáze: MEDLINE
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