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^*

^*Corresponding author for this work

AIMMS

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 (EC₅₀ 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	Humana Press Inc
Pages	505-520
Number of pages	16
ISBN (Electronic)	9781071620991
ISBN (Print)	9781071620984, 9781071621011
DOIs	https://doi.org/10.1007/978-1-0716-2099-1_23
Publication status	Published - 2022

Publication series

Name	Methods in Molecular Biology
Volume	2451
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

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.

Funding

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).

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

Persistent URL (handle)

Persistent link

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). Humana Press Inc. 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. Humana Press Inc, 2022. pp. 505-520 (Methods in Molecular Biology).

@inbook{e8e0a8432cc045e19c24d22dc286126e,

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

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 (kD in low nM range). Subsequently, upon illumination, these conjugates are very toxic and selective to cells overexpressing the target of interest (EC50 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.",

keywords = "Binding affinity, Co-cultures, Intracellular degradation, Nanobody-photosensitizer conjugate, Selective toxicity, Uptake",

author = "\{Beltr{\'a}n Hern{\'a}ndez\}, Irati and \{De Groof\}, \{Timo W.M.\} and Raimond Heukers and Sabrina Oliveira",

note = "Funding Information: The authors received funding from the European Research Council (ERC) under the European Union{\textquoteright}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: {\textcopyright} 2022, Springer Science+Business Media, LLC, part of Springer Nature.",

year = "2022",

doi = "10.1007/978-1-0716-2099-1\_23",

language = "English",

isbn = "9781071620984",

series = "Methods in Molecular Biology",

publisher = "Humana Press Inc",

pages = "505--520",

editor = "Mans Broekgaarden and Hong Zhang and Mladen Korbelik and Hamblin, \{Michael R.\} and Michal Heger",

booktitle = "Photodynamic Therapy",

}

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, Humana Press Inc, 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. Humana Press Inc, 2022. p. 505-520 (Methods in Molecular Biology; Vol. 2451).

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

TY - CHAP

T1 - In Vitro Assessment of Binding Affinity, Selectivity, Uptake, Intracellular Degradation, and Toxicity of Nanobody-Photosensitizer Conjugates

AU - Beltrán Hernández, Irati

AU - De Groof, Timo W.M.

AU - Heukers, Raimond

AU - Oliveira, Sabrina

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.

PY - 2022

Y1 - 2022

N2 - 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 (kD in low nM range). Subsequently, upon illumination, these conjugates are very toxic and selective to cells overexpressing the target of interest (EC50 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.

AB - 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 (kD in low nM range). Subsequently, upon illumination, these conjugates are very toxic and selective to cells overexpressing the target of interest (EC50 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.

KW - Binding affinity

KW - Co-cultures

KW - Intracellular degradation

KW - Nanobody-photosensitizer conjugate

KW - Selective toxicity

KW - Uptake

UR - https://www.scopus.com/pages/publications/85129629621

UR - https://www.scopus.com/inward/citedby.url?scp=85129629621&partnerID=8YFLogxK

UR - https://pubmed.ncbi.nlm.nih.gov/35505028/

U2 - 10.1007/978-1-0716-2099-1_23

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

M3 - Chapter

C2 - 35505028

AN - SCOPUS:85129629621

SN - 9781071620984

SN - 9781071621011

T3 - Methods in Molecular Biology

SP - 505

EP - 520

BT - Photodynamic Therapy

A2 - Broekgaarden, Mans

A2 - Zhang, Hong

A2 - Korbelik, Mladen

A2 - Hamblin, Michael R.

A2 - Heger, Michal

PB - Humana Press Inc

ER -

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. Humana Press Inc. 2022. p. 505-520. (Methods in Molecular Biology). Epub 2022 May 4. 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

Funding

Keywords

UN SDGs

Access to Document

Persistent URL (handle)

Other files and links

Fingerprint

Cite this