Academic Journal
Efficiency landscape of bioorthogonal click reactions producing bispecific antibody conjugates.
| Τίτλος: | Efficiency landscape of bioorthogonal click reactions producing bispecific antibody conjugates. |
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| Συγγραφείς: | Petri L; Medicinal Chemistry Research Group, HUN-REN Research Centre for Natural Sciences, 2 Magyar tudósok krt, 1117 Budapest, Hungary; National Drug Discovery and Development Laboratory, HUN-REN Research Centre for Natural Sciences, 2 Magyar tudósok krt, 1117 Budapest, Hungary., Kovács ÍK; Medicinal Chemistry Research Group, HUN-REN Research Centre for Natural Sciences, 2 Magyar tudósok krt, 1117 Budapest, Hungary; National Drug Discovery and Development Laboratory, HUN-REN Research Centre for Natural Sciences, 2 Magyar tudósok krt, 1117 Budapest, Hungary; Department of Organic Chemistry and Technology, Budapest University of Technology and Economics, 4 Szt. Gellért tér, 1111 Budapest, Hungary., Szabó B; Medicinal Chemistry Research Group, HUN-REN Research Centre for Natural Sciences, 2 Magyar tudósok krt, 1117 Budapest, Hungary; National Drug Discovery and Development Laboratory, HUN-REN Research Centre for Natural Sciences, 2 Magyar tudósok krt, 1117 Budapest, Hungary; Department of Organic Chemistry and Technology, Budapest University of Technology and Economics, 4 Szt. Gellért tér, 1111 Budapest, Hungary., Imre T; Medicinal Chemistry Research Group, HUN-REN Research Centre for Natural Sciences, 2 Magyar tudósok krt, 1117 Budapest, Hungary; National Drug Discovery and Development Laboratory, HUN-REN Research Centre for Natural Sciences, 2 Magyar tudósok krt, 1117 Budapest, Hungary; MS Metabolomics Research Laboratory, HUN-REN Research Centre for Natural Sciences, 2 Magyar tudósok krt, 1117 Budapest, Hungary., Rácz A; National Drug Discovery and Development Laboratory, HUN-REN Research Centre for Natural Sciences, 2 Magyar tudósok krt, 1117 Budapest, Hungary; Plasma Chemistry Research Group, HUN-REN Research Centre for Natural Sciences, 2 Magyar tudósok krt, 1117 Budapest, Hungary., Chudasama V; Department of Chemistry, University College London, 20 Gordon Street, London WC1H OAJ, UK., Szijj PA; Department of Chemistry, University College London, 20 Gordon Street, London WC1H OAJ, UK; Stanford Sarafan ChEM-H and Department of Chemistry, Stanford University, Stanford, CA 94305, USA. Electronic address: pszijj@stanford.edu., Keserű GM; Medicinal Chemistry Research Group, HUN-REN Research Centre for Natural Sciences, 2 Magyar tudósok krt, 1117 Budapest, Hungary; National Drug Discovery and Development Laboratory, HUN-REN Research Centre for Natural Sciences, 2 Magyar tudósok krt, 1117 Budapest, Hungary; Department of Organic Chemistry and Technology, Budapest University of Technology and Economics, 4 Szt. Gellért tér, 1111 Budapest, Hungary. Electronic address: keseru.gyorgy@ttk.hu. |
| Πηγή: | Cell reports methods [Cell Rep Methods] 2026 Jun 15; Vol. 6 (6), pp. 101416. Date of Electronic Publication: 2026 Apr 21. |
| Τύπος έκδοσης: | Journal Article |
| Γλώσσα: | English |
| Στοιχεία περιοδικού: | Publisher: Elsevier Inc Country of Publication: United States NLM ID: 9918227360606676 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2667-2375 (Electronic) Linking ISSN: 26672375 NLM ISO Abbreviation: Cell Rep Methods Subsets: MEDLINE |
| Imprint Name(s): | Original Publication: [New York] : Elsevier Inc., [2021]- |
| Ιατρικοί όροι (MeSH): | Antibodies, Bispecific*/chemistry , Click Chemistry*/methods , Immunoconjugates*/chemistry, Cycloaddition Reaction/methods ; Cyclooctanes/chemistry ; Humans |
| Περίληψη: | Bioorthogonal conjugation techniques offer a powerful and flexible approach for the modular construction of multifunctional biomolecules, such as bispecific antibodies. In this study, we systematically compared two inverse electron demand Diels-Alder (IEDDA) reactions, between tetrazine and either trans-cyclooctene (TCO) or bicyclo[6.1.0]nonyne (BCN), to generate chemically conjugated bispecific antibody constructs. We applied a design of experiments (DoE) framework to explore how various reaction parameters influence conjugation efficiency. The two systems exhibited distinct reactivity patterns: the BCN-tetrazine reaction proved to be more robust, while the TCO-tetrazine ligation showed a more complex dependency on reaction time and temperature. To assess biological functionality, the bispecific constructs were evaluated by ELISA, confirming preserved antigen-recognition after both conjugation strategies. Both strategies consistently yielded bispecific constructs with comparable physicochemical and functional profiles. These insights support the application of this chemical conjugation strategy as a rapid, tunable, and modular platform for early-stage multispecific antibody development. (Copyright © 2026 The Authors. Published by Elsevier Inc. All rights reserved.) |
| Competing Interests: | Declaration of interests The authors declare no competing interests. |
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| Contributed Indexing: | Keywords: CP: biotechnology; antibody; bioconjugation; bioorthogonal chemistry; bispecific antibody; click chemistry; reaction optimization |
| Substance Nomenclature: | 0 (Antibodies, Bispecific) 0 (Immunoconjugates) 0 (Cyclooctanes) |
| Entry Date(s): | Date Created: 20260422 Date Completed: 20260615 Latest Revision: 20260813 |
| Update Code: | 20260813 |
| PubMed Central ID: | PMC13282659 |
| DOI: | 10.1016/j.crmeth.2026.101416 |
| PMID: | 42019503 |
| Βάση Δεδομένων: | MEDLINE |
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