A Reagent for Amine-Directed Conjugation to IgG1 Antibodies

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A Reagent for Amine-Directed Conjugation to IgG1 Antibodies. / Märcher, Anders; Palmfeldt, Johan; Nisavic, Marija; Gothelf, Kurt Vesterager.

In: Angewandte Chemie International Edition, Vol. 60, No. 12, 03.2021, p. 6539-6544.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

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Märcher, A, Palmfeldt, J, Nisavic, M & Gothelf, KV 2021, 'A Reagent for Amine-Directed Conjugation to IgG1 Antibodies', Angewandte Chemie International Edition, vol. 60, no. 12, pp. 6539-6544. https://doi.org/10.1002/anie.202013911

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Märcher, Anders ; Palmfeldt, Johan ; Nisavic, Marija ; Gothelf, Kurt Vesterager. / A Reagent for Amine-Directed Conjugation to IgG1 Antibodies. In: Angewandte Chemie International Edition. 2021 ; Vol. 60, No. 12. pp. 6539-6544.

Bibtex

@article{3d28aca91f6c4a95bc56881871627c84,
title = "A Reagent for Amine-Directed Conjugation to IgG1 Antibodies",
abstract = "Functionalized antibodies are an indispensable resource for diagnosis, therapy and as a research tool for chemical biology. However, simpler and better methodologies are often required to improve the labeling of antibodies in terms of selectivity and scalability, in particular for the production of antibody drug conjugates. Herein, we report the development of an easily available chemical reagent that allows site-directed labeling of native human IgG1 antibodies. The labeling procedure has low time demand, and only one reagent is needed. The salicylaldehyde moiety of the reagent reacts with surface exposed lysine residues to transiently form an iminium ion, and this positions a semi-reactive ester into proximity of a second lysine residue that reacts with the ester to form an amide. Interestingly, it appears that the formation of the iminium ion also has a significant activating effect of the ester. We show that the reagent can effectively be used to conjugate functionalities of interest to human IgG1 antibodies in good yield and conversion. Moreover, the reagent allows for high control of number of labels per antibody. We use flow cytometry and bio-layer interferometry to confirm that the labeled antibodies retain antigen binding.",
keywords = "IgG1, antibodies, fluorophore, labeling, site-direction",
author = "Anders M{\"a}rcher and Johan Palmfeldt and Marija Nisavic and Gothelf, {Kurt Vesterager}",
note = "{\textcopyright} 2020 Wiley-VCH GmbH.",
year = "2021",
month = mar,
doi = "10.1002/anie.202013911",
language = "English",
volume = "60",
pages = "6539--6544",
journal = "Angewandte Chemie International Edition",
issn = "1433-7851",
publisher = "Wiley - VCH Verlag GmbH & CO. KGaA",
number = "12",

}

RIS

TY - JOUR

T1 - A Reagent for Amine-Directed Conjugation to IgG1 Antibodies

AU - Märcher, Anders

AU - Palmfeldt, Johan

AU - Nisavic, Marija

AU - Gothelf, Kurt Vesterager

N1 - © 2020 Wiley-VCH GmbH.

PY - 2021/3

Y1 - 2021/3

N2 - Functionalized antibodies are an indispensable resource for diagnosis, therapy and as a research tool for chemical biology. However, simpler and better methodologies are often required to improve the labeling of antibodies in terms of selectivity and scalability, in particular for the production of antibody drug conjugates. Herein, we report the development of an easily available chemical reagent that allows site-directed labeling of native human IgG1 antibodies. The labeling procedure has low time demand, and only one reagent is needed. The salicylaldehyde moiety of the reagent reacts with surface exposed lysine residues to transiently form an iminium ion, and this positions a semi-reactive ester into proximity of a second lysine residue that reacts with the ester to form an amide. Interestingly, it appears that the formation of the iminium ion also has a significant activating effect of the ester. We show that the reagent can effectively be used to conjugate functionalities of interest to human IgG1 antibodies in good yield and conversion. Moreover, the reagent allows for high control of number of labels per antibody. We use flow cytometry and bio-layer interferometry to confirm that the labeled antibodies retain antigen binding.

AB - Functionalized antibodies are an indispensable resource for diagnosis, therapy and as a research tool for chemical biology. However, simpler and better methodologies are often required to improve the labeling of antibodies in terms of selectivity and scalability, in particular for the production of antibody drug conjugates. Herein, we report the development of an easily available chemical reagent that allows site-directed labeling of native human IgG1 antibodies. The labeling procedure has low time demand, and only one reagent is needed. The salicylaldehyde moiety of the reagent reacts with surface exposed lysine residues to transiently form an iminium ion, and this positions a semi-reactive ester into proximity of a second lysine residue that reacts with the ester to form an amide. Interestingly, it appears that the formation of the iminium ion also has a significant activating effect of the ester. We show that the reagent can effectively be used to conjugate functionalities of interest to human IgG1 antibodies in good yield and conversion. Moreover, the reagent allows for high control of number of labels per antibody. We use flow cytometry and bio-layer interferometry to confirm that the labeled antibodies retain antigen binding.

KW - IgG1

KW - antibodies

KW - fluorophore

KW - labeling

KW - site-direction

UR - http://www.scopus.com/inward/record.url?scp=85100343789&partnerID=8YFLogxK

U2 - 10.1002/anie.202013911

DO - 10.1002/anie.202013911

M3 - Journal article

C2 - 33306206

VL - 60

SP - 6539

EP - 6544

JO - Angewandte Chemie International Edition

JF - Angewandte Chemie International Edition

SN - 1433-7851

IS - 12

ER -