Bioconjugation of a Near-Infrared DNA-Stabilized Silver Nanocluster to Peptides and Human Insulin by Copper-Free Click Chemistry

Vanessa Rück, Narendra K. Mishra, Kasper K. Sørensen, Mikkel B. Liisberg, Ane B. Sloth, Cecilia Cerretani, Christian B. Mollerup, Andreas Kjaer, Chenguang Lou, Knud J. Jensen, Tom Vosch*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

13 Citations (Scopus)

Abstract

DNA-stabilized silver nanoclusters (DNA-AgNCs) are biocompatible emitters with intriguing properties. However, they have not been extensively used for bioimaging applications due to the lack of structural information and hence predictable conjugation strategies. Here, a copper-free click chemistry method for linking a well-characterized DNA-AgNC to molecules of interest is presented. Three different peptides and a small protein, human insulin, were tested as labeling targets. The conjugation to the target compounds was verified by MS, HPLC, and time-resolved anisotropy measurements. Moreover, the spectroscopic properties of DNA-AgNCs were found to be unaffected by the linking reactions. For DNA-AgNC-conjugated human insulin, fluorescence imaging studies were performed on Chinese hamster ovary (CHO) cells overexpressing human insulin receptor B (hIR-B). The specific staining of the CHO cell membranes demonstrates that DNA-AgNCs are great candidates for bioimaging applications, and the proposed linking strategy is easy to implement when the DNA-AgNC structure is known.

Original languageEnglish
JournalJournal of the American Chemical Society
Volume145
Issue number30
Pages (from-to)16771–16777
Number of pages7
ISSN0002-7863
DOIs
Publication statusPublished - 2023

Bibliographical note

Funding Information:
V.R., M.B.L., C.C., and T.V. acknowledge funding from the Villum Foundation (VKR023115) and the Independent Research Fund Denmark (0136-00024B). N.K.M., K.K.S., and K.J.J. thank the Villum Foundation (VKR18333) for funding the Biomolecular Nanoscale Engineering Center (BioNEC), a VILLUM center of excellence, and the Novo Nordisk Foundation funding for the Center for Biopharmaceuticals and Biobarriers in Drug Delivery (BioDelivery; Grand Challenge Program; NNF16OC0021948). The authors would like to thank Morten Lundh from Gubra for providing the CHO cells expressing the hIR-B.

Publisher Copyright:
© 2023 The Authors. Published by American Chemical Society.

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