Oriented conjugation of luciferase to infrared quantum dots enables BRET-based enzyme sensing

Padric M. Garden; Allison M. Dennis

doi:10.1117/12.2608887

3 March 2022 Oriented conjugation of luciferase to infrared quantum dots enables BRET-based enzyme sensing

Padric M. Garden, Allison M. Dennis

Proceedings Volume PC11977, Colloidal Nanoparticles for Biomedical Applications XVII; PC1197708 (2022) https://doi.org/10.1117/12.2608887
Event: SPIE BiOS, 2022, San Francisco, California, United States

Abstract

Luciferase-quantum dot (QD) conjugates enable excitation-free in vitro and in vivo imaging, but their use as sensors has been limited due to the use of randomly oriented conjugation strategies. We have developed an N-terminal specific conjugation strategy to orient NanoLuc luciferase on zwitterionic polymer-coated infrared QDs using a recently demonstrated 6-(azidomethyl)-2-pyridinecarbaldehyde-mediated azide labeling reaction. Using this strategy, we insert enzyme substrate peptide sequences between the luciferase and QD enabling detection of cleavage enzyme activity through bioluminescence resonance energy transfer (BRET). We optimized and characterized these sensor constructs and demonstrate their ability to access enzyme activity.

Conference Presentation

Citation Download Citation

Padric M. Garden and Allison M. Dennis "Oriented conjugation of luciferase to infrared quantum dots enables BRET-based enzyme sensing", Proc. SPIE PC11977, Colloidal Nanoparticles for Biomedical Applications XVII, PC1197708 (3 March 2022); https://doi.org/10.1117/12.2608887

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