Skip to main content

Lund University Publications

LUND UNIVERSITY LIBRARIES

Impact of site-specific conjugation strategies on the pharmacokinetics of antibody conjugated radiotherapeutics

Nagy, Ábel ; Ulmert, David LU ; Zedan, Wahed LU ; Storey, Claire M. ; Park, Julie ; Geres, Susanne LU ; Lückerath, Katharina ; Sjöström, Kjell ; Westin, Hadis and Peekhaus, Norbert , et al. (2024) In European Journal of Medicinal Chemistry 280.
Abstract

Antibody radionuclide conjugates are an emerging modality for targeted imaging and potent therapy of disseminated disease. Coupling of radionuclides to monoclonal antibodies (mAbs) is typically achieved by applying non-site-specific labelling techniques. With the ambition of reducing variability, increasing labelling efficacy and stability, several site-specific conjugation strategies have been developed in recent years for toxin- and fluorophore-mAb conjugates. In this study, we studied two site-specific labelling strategies for the conjugation of the macrocyclic chelating agent, DOTA, to the anti-Leucine Rich Repeat Containing 15 (LRRC15) mAb DUNP19. Specifically, one approach utilized a DOTA-bearing peptide (FcIII) with a strong... (More)

Antibody radionuclide conjugates are an emerging modality for targeted imaging and potent therapy of disseminated disease. Coupling of radionuclides to monoclonal antibodies (mAbs) is typically achieved by applying non-site-specific labelling techniques. With the ambition of reducing variability, increasing labelling efficacy and stability, several site-specific conjugation strategies have been developed in recent years for toxin- and fluorophore-mAb conjugates. In this study, we studied two site-specific labelling strategies for the conjugation of the macrocyclic chelating agent, DOTA, to the anti-Leucine Rich Repeat Containing 15 (LRRC15) mAb DUNP19. Specifically, one approach utilized a DOTA-bearing peptide (FcIII) with a strong affinity for the fragment crystallizable (Fc) domain of the human IgG1 of DUNP19 (DUNP19LF-FcIII-DOTASS), while the other leveraged a chemo-enzymatic technique to substitute the N-linked bi-antennary oligosaccharides in the human IgG1 Fc domain with DOTA (DUNP19LF-gly-DOTASS). To assess if these methods impact the antibody's binding properties and targeting efficacy, comparative in vitro and in vivo studies of the generated DUNP19-conjugates were performed. While the LRRC15 binding of both radioimmunoconjugates remained intact, the conjugation methods had different impacts on their abilities to interact with FcRn and FcγRs. In vitro assessments of DUNP19LF-FcIII-DOTASS and DUNP19LF-gly-DOTASS demonstrated markedly decreased affinity for FcRn and FcγRIIIa (CD16), respectively. DUNP19LF-FcIII-DOTASS demonstrated increased blood and tissue kinetics in vivo, confirming loss of FcRn binding. While the ablated FcγR interaction of DUNP19LF-gly-DOTASS had no immediate impact on in vivo biodistribution, reduced immunotherapeutic effect can be expected in future studies as a result of reduced NK-cells interaction. In conclusion, our findings underscore the necessity for meticulous consideration and evaluation of mAb labelling strategies, extending beyond mere conjugation efficiency and radiolabeling yields. Notably, site-specific labelling methods were found to significantly influence the immunological impact of Fc interactions. Therefore, it is of paramount importance to consider the intended diagnostic or therapeutic application of the construct and to adopt conjugation strategies that ensure the preservation of critical pharmacological properties and functionality of the antibody in use.

(Less)
Please use this url to cite or link to this publication:
author
; ; ; ; ; ; ; ; and , et al. (More)
; ; ; ; ; ; ; ; ; ; ; and (Less)
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Antibody labelling, FcIII peptide, Glycan engineering, Radioimmunotherapy, Site-specific labelling
in
European Journal of Medicinal Chemistry
volume
280
article number
116927
publisher
Elsevier Masson SAS
external identifiers
  • scopus:85205534815
  • pmid:39378827
ISSN
0223-5234
DOI
10.1016/j.ejmech.2024.116927
language
English
LU publication?
yes
additional info
Publisher Copyright: © 2024 The Authors
id
ea3871fd-4a1f-452a-b192-65541af055fd
date added to LUP
2024-11-27 15:14:26
date last changed
2024-12-11 16:54:41
@article{ea3871fd-4a1f-452a-b192-65541af055fd,
  abstract     = {{<p>Antibody radionuclide conjugates are an emerging modality for targeted imaging and potent therapy of disseminated disease. Coupling of radionuclides to monoclonal antibodies (mAbs) is typically achieved by applying non-site-specific labelling techniques. With the ambition of reducing variability, increasing labelling efficacy and stability, several site-specific conjugation strategies have been developed in recent years for toxin- and fluorophore-mAb conjugates. In this study, we studied two site-specific labelling strategies for the conjugation of the macrocyclic chelating agent, DOTA, to the anti-Leucine Rich Repeat Containing 15 (LRRC15) mAb DUNP19. Specifically, one approach utilized a DOTA-bearing peptide (FcIII) with a strong affinity for the fragment crystallizable (Fc) domain of the human IgG<sub>1</sub> of DUNP19 (DUNP19<sup>LF</sup>-FcIII-DOTA<sup>SS</sup>), while the other leveraged a chemo-enzymatic technique to substitute the N-linked bi-antennary oligosaccharides in the human IgG<sub>1</sub> Fc domain with DOTA (DUNP19<sup>LF</sup>-gly-DOTA<sup>SS</sup>). To assess if these methods impact the antibody's binding properties and targeting efficacy, comparative in vitro and in vivo studies of the generated DUNP19-conjugates were performed. While the LRRC15 binding of both radioimmunoconjugates remained intact, the conjugation methods had different impacts on their abilities to interact with FcRn and FcγRs. In vitro assessments of DUNP19<sup>LF</sup>-FcIII-DOTA<sup>SS</sup> and DUNP19<sup>LF</sup>-gly-DOTA<sup>SS</sup> demonstrated markedly decreased affinity for FcRn and FcγRIIIa (CD16), respectively. DUNP19<sup>LF</sup>-FcIII-DOTA<sup>SS</sup> demonstrated increased blood and tissue kinetics in vivo, confirming loss of FcRn binding. While the ablated FcγR interaction of DUNP19<sup>LF</sup>-gly-DOTA<sup>SS</sup> had no immediate impact on in vivo biodistribution, reduced immunotherapeutic effect can be expected in future studies as a result of reduced NK-cells interaction. In conclusion, our findings underscore the necessity for meticulous consideration and evaluation of mAb labelling strategies, extending beyond mere conjugation efficiency and radiolabeling yields. Notably, site-specific labelling methods were found to significantly influence the immunological impact of Fc interactions. Therefore, it is of paramount importance to consider the intended diagnostic or therapeutic application of the construct and to adopt conjugation strategies that ensure the preservation of critical pharmacological properties and functionality of the antibody in use.</p>}},
  author       = {{Nagy, Ábel and Ulmert, David and Zedan, Wahed and Storey, Claire M. and Park, Julie and Geres, Susanne and Lückerath, Katharina and Sjöström, Kjell and Westin, Hadis and Peekhaus, Norbert and Thorek, Daniel LJ and Karlström, Amelie Eriksson and Altai, Mohamed}},
  issn         = {{0223-5234}},
  keywords     = {{Antibody labelling; FcIII peptide; Glycan engineering; Radioimmunotherapy; Site-specific labelling}},
  language     = {{eng}},
  month        = {{12}},
  publisher    = {{Elsevier Masson SAS}},
  series       = {{European Journal of Medicinal Chemistry}},
  title        = {{Impact of site-specific conjugation strategies on the pharmacokinetics of antibody conjugated radiotherapeutics}},
  url          = {{http://dx.doi.org/10.1016/j.ejmech.2024.116927}},
  doi          = {{10.1016/j.ejmech.2024.116927}},
  volume       = {{280}},
  year         = {{2024}},
}