In Vitro Assessment of Binding Affinity, Selectivity, Uptake, Intracellular Degradation, and Toxicity of Nanobody-Photosensitizer Conjugates

Irati Beltrán Hernández; Timo W M De Groof; Raimond Heukers; Sabrina Oliveira

doi:10.1007/978-1-0716-2099-1_23

In Vitro Assessment of Binding Affinity, Selectivity, Uptake, Intracellular Degradation, and Toxicity of Nanobody-Photosensitizer Conjugates

Irati Beltrán Hernández, Timo W M De Groof, Raimond Heukers, Sabrina Oliveira

VU University Medical Center Amsterdam

Research output: Chapter in Book/Report/Conference proceeding › Chapter › Academic › peer-review

Abstract

Photosensitizers have recently been conjugated to nanobodies for targeted photodynamic therapy (PDT) to selectively kill cancer cells. The success of this approach relies on nanobody-photosensitizer conjugates that bind specifically to their targets with very high affinities (k D in low nM range). Subsequently, upon illumination, these conjugates are very toxic and selective to cells overexpressing the target of interest (EC 50 in low nM range). In this chapter, protocols are described to determine the binding affinity of the nanobody-photosensitizer conjugates and assess the toxicity and selectivity of the conjugates when performing in vitro PDT studies. In addition, and because the efficacy of PDT also depends on the (subcellular) localization of the conjugates at the time of illumination, assays are described to investigate the uptake and the intracellular degradation of the nanobody-photosensitizer conjugates.

Original language	English
Title of host publication	Photodynamic Therapy
Subtitle of host publication	Methods and Protocols
Editors	Mans Broekgaarden, Hong Zhang, Mladen Korbelik, Michael R. Hamblin, Michal Heger
Publisher	Springer
Chapter	23
Pages	505-520
Number of pages	16
ISBN (Electronic)	978-1-0716-2099-1
ISBN (Print)	978-1-0716-2098-4
DOIs	https://doi.org/10.1007/978-1-0716-2099-1_23
Publication status	Published - 2022

Publication series

Name	Methods in Molecular Biology
Publisher	Humana Press
Volume	2451
ISSN (Print)	1064-3745

Bibliographical note

Funding Information:
The authors received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program (grant agreement No. 677582) and the Netherlands Organization for Scientific Research (NWO: Vici grant 016.140.657).

Publisher Copyright:
© 2022, Springer Science+Business Media, LLC, part of Springer Nature.

Keywords

Binding affinity
Co-cultures
Intracellular degradation
Nanobody-photosensitizer conjugate
Selective toxicity
Uptake

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1007/978-1-0716-2099-1_23

Pages from 978-1-0716-2099-1bFinal published version, 289 KBLicence: Taverne

Cite this

Beltrán Hernández, I., De Groof, T. W. M., Heukers, R., & Oliveira, S. (2022). In Vitro Assessment of Binding Affinity, Selectivity, Uptake, Intracellular Degradation, and Toxicity of Nanobody-Photosensitizer Conjugates. In M. Broekgaarden, H. Zhang, M. Korbelik, M. R. Hamblin, & M. Heger (Eds.), Photodynamic Therapy: Methods and Protocols (pp. 505-520). (Methods in Molecular Biology; Vol. 2451). Springer. https://doi.org/10.1007/978-1-0716-2099-1_23

Beltrán Hernández, Irati ; De Groof, Timo W M ; Heukers, Raimond et al. / In Vitro Assessment of Binding Affinity, Selectivity, Uptake, Intracellular Degradation, and Toxicity of Nanobody-Photosensitizer Conjugates. Photodynamic Therapy: Methods and Protocols. editor / Mans Broekgaarden ; Hong Zhang ; Mladen Korbelik ; Michael R. Hamblin ; Michal Heger. Springer, 2022. pp. 505-520 (Methods in Molecular Biology).

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Beltrán Hernández, I, De Groof, TWM, Heukers, R & Oliveira, S 2022, In Vitro Assessment of Binding Affinity, Selectivity, Uptake, Intracellular Degradation, and Toxicity of Nanobody-Photosensitizer Conjugates. in M Broekgaarden, H Zhang, M Korbelik, MR Hamblin & M Heger (eds), Photodynamic Therapy: Methods and Protocols. Methods in Molecular Biology, vol. 2451, Springer, pp. 505-520. https://doi.org/10.1007/978-1-0716-2099-1_23

In Vitro Assessment of Binding Affinity, Selectivity, Uptake, Intracellular Degradation, and Toxicity of Nanobody-Photosensitizer Conjugates. / Beltrán Hernández, Irati; De Groof, Timo W M; Heukers, Raimond et al.
Photodynamic Therapy: Methods and Protocols. ed. / Mans Broekgaarden; Hong Zhang; Mladen Korbelik; Michael R. Hamblin; Michal Heger. Springer, 2022. p. 505-520 (Methods in Molecular Biology; Vol. 2451).

Research output: Chapter in Book/Report/Conference proceeding › Chapter › Academic › peer-review

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AU - Heukers, Raimond

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N1 - Funding Information: The authors received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program (grant agreement No. 677582) and the Netherlands Organization for Scientific Research (NWO: Vici grant 016.140.657). Publisher Copyright: © 2022, Springer Science+Business Media, LLC, part of Springer Nature.

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Beltrán Hernández I, De Groof TWM, Heukers R, Oliveira S. In Vitro Assessment of Binding Affinity, Selectivity, Uptake, Intracellular Degradation, and Toxicity of Nanobody-Photosensitizer Conjugates. In Broekgaarden M, Zhang H, Korbelik M, Hamblin MR, Heger M, editors, Photodynamic Therapy: Methods and Protocols. Springer. 2022. p. 505-520. (Methods in Molecular Biology). doi: 10.1007/978-1-0716-2099-1_23

In Vitro Assessment of Binding Affinity, Selectivity, Uptake, Intracellular Degradation, and Toxicity of Nanobody-Photosensitizer Conjugates

Abstract

Publication series

Bibliographical note

Keywords

UN SDGs

Access to Document

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