Comparison of NIR FRET pairs for quantitative transferrin-based assay

Nattawut Sinsuebphon; Travis Bevington; Lingling Zhao; Abe Ken; Margarida Barroso; Xavier Intes

doi:10.1117/12.2040097

17 February 2014 Comparison of NIR FRET pairs for quantitative transferrin-based assay

Nattawut Sinsuebphon, Travis Bevington, Lingling Zhao, Abe Ken, Margarida Barroso, Xavier Intes

Proceedings Volume 8937, Multimodal Biomedical Imaging IX; 89370X (2014) https://doi.org/10.1117/12.2040097
Event: SPIE BiOS, 2014, San Francisco, California, United States

Abstract

Transferrin (Tfn) is commonly used as a drug delivery carrier for cancer treatment. Tfn cellular internalization can be observed by Förster resonance energy transfer (FRET), which occurs when two fluorophores - donor and acceptor - are a few nanometers apart. Donor fluorescence lifetime can be used to sense and quantify FRET occurrence. In FRET state, the donor is quenched leading to a significant reduction in its lifetime. In this study, donor and acceptor near-infrared (NIR) fluorophore-labeled Tfn were used to quantify cellular internalization in breast cancer cell line (T47D). Based on donor lifetime, quantum yield and spectral data, seven NIR FRET pairs were chosen for this comparison. Performance of the different NIR FRET pairs was evaluated in vitro in multiwell plate settings and by analyzing the relationship between quenched donor fraction and acceptor:donor ratio. Additionally, we performed brightness comparison between each pairs. Several parameters, such as brightness, lifetime, R0 and FRET donor population values are used to identify the most suitable NIR FRET pair for in vivo studies in preclinical settings.

Citation Download Citation

Nattawut Sinsuebphon, Travis Bevington, Lingling Zhao, Abe Ken, Margarida Barroso, and Xavier Intes "Comparison of NIR FRET pairs for quantitative transferrin-based assay", Proc. SPIE 8937, Multimodal Biomedical Imaging IX, 89370X (17 February 2014); https://doi.org/10.1117/12.2040097

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