Introduction of an Aldehyde Handle on Nanobodies by Affinity-Guided Labeling

Michael R. Mortensen; Mikkel B. Skovsgaard; Anders Märcher; Veronica L. Andersen; Johan Palmfeldt; Thorbjørn B. Nielsen; Thomas Tørring; Nick S. Laursen; Kasper R. Andersen; Jørgen Kjems; Kurt V. Gothelf

doi:10.1021/acs.bioconjchem.0c00151

Introduction of an Aldehyde Handle on Nanobodies by Affinity-Guided Labeling

Michael R. Mortensen, Mikkel B. Skovsgaard, Anders Märcher, Veronica L. Andersen, Johan Palmfeldt, Thorbjørn B. Nielsen, Thomas Tørring, Nick S. Laursen, Kasper R. Andersen, Jørgen Kjems, Kurt V. Gothelf

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

9 Citations (Scopus)

Abstract

Chemically modified antigen-binding proteins are widely applied for their targeting abilities in the fields of biotechnology, medicine, and diagnostics. However, the production of site-selectively modified proteins remains a challenge. Here, we have designed a chemical probe for the introduction of a reactive aldehyde on nanobodies by metal-complex-guided conjugation. The probe design allows for purification of the conjugates, and the aldehyde constitutes an efficient handle for further modification of the nanobodies. In vitro experiments confirmed the binding activity and selectivity of fluorescent conjugates toward the native antigen. Furthermore, the modification strategy allowed for production of a nanobody-drug conjugate that was active in vitro.

Original language	English
Journal	Bioconjugate Chemistry
Volume	31
Issue	5
Pages (from-to)	1295-1300
Number of pages	6
ISSN	1043-1802
DOIs	https://doi.org/10.1021/acs.bioconjchem.0c00151
Publication status	Published - May 2020

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title = "Introduction of an Aldehyde Handle on Nanobodies by Affinity-Guided Labeling",

abstract = "Chemically modified antigen-binding proteins are widely applied for their targeting abilities in the fields of biotechnology, medicine, and diagnostics. However, the production of site-selectively modified proteins remains a challenge. Here, we have designed a chemical probe for the introduction of a reactive aldehyde on nanobodies by metal-complex-guided conjugation. The probe design allows for purification of the conjugates, and the aldehyde constitutes an efficient handle for further modification of the nanobodies. In vitro experiments confirmed the binding activity and selectivity of fluorescent conjugates toward the native antigen. Furthermore, the modification strategy allowed for production of a nanobody-drug conjugate that was active in vitro.",

author = "Mortensen, {Michael R.} and Skovsgaard, {Mikkel B.} and Anders M{\"a}rcher and Andersen, {Veronica L.} and Johan Palmfeldt and Nielsen, {Thorbj{\o}rn B.} and Thomas T{\o}rring and Laursen, {Nick S.} and Andersen, {Kasper R.} and J{\o}rgen Kjems and Gothelf, {Kurt V.}",

year = "2020",

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doi = "10.1021/acs.bioconjchem.0c00151",

language = "English",

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T1 - Introduction of an Aldehyde Handle on Nanobodies by Affinity-Guided Labeling

AU - Mortensen, Michael R.

AU - Skovsgaard, Mikkel B.

AU - Märcher, Anders

AU - Andersen, Veronica L.

AU - Palmfeldt, Johan

AU - Nielsen, Thorbjørn B.

AU - Tørring, Thomas

AU - Laursen, Nick S.

AU - Andersen, Kasper R.

AU - Kjems, Jørgen

AU - Gothelf, Kurt V.

PY - 2020/5

Y1 - 2020/5

N2 - Chemically modified antigen-binding proteins are widely applied for their targeting abilities in the fields of biotechnology, medicine, and diagnostics. However, the production of site-selectively modified proteins remains a challenge. Here, we have designed a chemical probe for the introduction of a reactive aldehyde on nanobodies by metal-complex-guided conjugation. The probe design allows for purification of the conjugates, and the aldehyde constitutes an efficient handle for further modification of the nanobodies. In vitro experiments confirmed the binding activity and selectivity of fluorescent conjugates toward the native antigen. Furthermore, the modification strategy allowed for production of a nanobody-drug conjugate that was active in vitro.

AB - Chemically modified antigen-binding proteins are widely applied for their targeting abilities in the fields of biotechnology, medicine, and diagnostics. However, the production of site-selectively modified proteins remains a challenge. Here, we have designed a chemical probe for the introduction of a reactive aldehyde on nanobodies by metal-complex-guided conjugation. The probe design allows for purification of the conjugates, and the aldehyde constitutes an efficient handle for further modification of the nanobodies. In vitro experiments confirmed the binding activity and selectivity of fluorescent conjugates toward the native antigen. Furthermore, the modification strategy allowed for production of a nanobody-drug conjugate that was active in vitro.

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U2 - 10.1021/acs.bioconjchem.0c00151

DO - 10.1021/acs.bioconjchem.0c00151

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Introduction of an Aldehyde Handle on Nanobodies by Affinity-Guided Labeling

Abstract

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