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

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

9 Citationer (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.

Originalsprog	Engelsk
Tidsskrift	Bioconjugate Chemistry
Vol/bind	31
Nummer	5
Sider (fra-til)	1295-1300
Antal sider	6
ISSN	1043-1802
DOI	https://doi.org/10.1021/acs.bioconjchem.0c00151
Status	Udgivet - maj 2020

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Citationsformater

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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",

month = may,

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

language = "English",

volume = "31",

pages = "1295--1300",

journal = "Bioconjugate Chemistry",

issn = "1043-1802",

publisher = "American Chemical Society",

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TY - JOUR

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.

UR - http://www.scopus.com/inward/record.url?scp=85085265715&partnerID=8YFLogxK

U2 - 10.1021/acs.bioconjchem.0c00151

DO - 10.1021/acs.bioconjchem.0c00151

M3 - Journal article

C2 - 32320218

AN - SCOPUS:85085265715

SN - 1043-1802

VL - 31

SP - 1295

EP - 1300

JO - Bioconjugate Chemistry

JF - Bioconjugate Chemistry

IS - 5

ER -

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

Abstract

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