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Tytuł:
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Small p53 derived peptide suitable for robust nanobodies dimerization.
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Autorzy:
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Dietsch F; Université de Strasbourg, UMR7242 Biotechnologie et Signalisation Cellulaire, Ecole Supérieure de Biotechnologie Strasbourg, F-67412 Illkirch, France.
Nominé Y; Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch, F-67412 Illkirch, France.
Stoessel A; Université de Strasbourg, UMR7242 Biotechnologie et Signalisation Cellulaire, Ecole Supérieure de Biotechnologie Strasbourg, F-67412 Illkirch, France.
Kostmann C; Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch, F-67412 Illkirch, France.
Bonhoure A; Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch, F-67412 Illkirch, France.
Chatton B; Université de Strasbourg, UMR7242 Biotechnologie et Signalisation Cellulaire, Ecole Supérieure de Biotechnologie Strasbourg, F-67412 Illkirch, France.
Donzeau M; Université de Strasbourg, UMR7242 Biotechnologie et Signalisation Cellulaire, Ecole Supérieure de Biotechnologie Strasbourg, F-67412 Illkirch, France. Electronic address: .
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Źródło:
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Journal of immunological methods [J Immunol Methods] 2021 Nov; Vol. 498, pp. 113144. Date of Electronic Publication: 2021 Sep 03.
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Typ publikacji:
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Journal Article; Research Support, Non-U.S. Gov't
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Język:
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English
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Imprint Name(s):
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Publication: Amsterdam : Elsevier
Original Publication: Amsterdam, North-Holand,
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MeSH Terms:
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Epitopes*
Green Fluorescent Proteins/*immunology
Peptide Fragments/*immunology
Single-Domain Antibodies/*immunology
Tumor Suppressor Protein p53/*immunology
Animals ; Antibody Affinity ; Antibody Specificity ; Green Fluorescent Proteins/genetics ; HeLa Cells ; Humans ; Mutation ; Peptide Fragments/genetics ; Protein Multimerization ; Tumor Suppressor Protein p53/genetics
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Contributed Indexing:
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Keywords: Avidity; Bivalency; Dimerization domain; Peptide; V(H)Hs; p53
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Substance Nomenclature:
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0 (Epitopes)
0 (Peptide Fragments)
0 (Single-Domain Antibodies)
0 (Tumor Suppressor Protein p53)
0 (enhanced green fluorescent protein)
147336-22-9 (Green Fluorescent Proteins)
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Entry Date(s):
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Date Created: 20210905 Date Completed: 20211108 Latest Revision: 20211108
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Update Code:
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20240105
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DOI:
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10.1016/j.jim.2021.113144
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PMID:
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34481824
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Bivalent V H Hs have been shown to display better functional affinity compared with their monovalent counterparts. Bivalency can be achieved either by inserting a hinge region between both V H Hs units or by using modules that lead to dimerization. In this report, a small self-associating peptide originating from the tetramerization domain of p53 was developed as a tool for devicing nanobody dimerization. This E3 peptide was evaluated for the dimerization of an anti-eGFP nanobody (nano-eGFP-E3) whose activity was compared to a bivalent anti-eGFP constructed in tandem using GS rich linker. The benefit of bivalency in terms of avidity and specificity was assessed in different in vitro and in cellulo assays. In ELISA and SPR, the dimeric and tandem formats were nearly equivalent in terms of gain of avidity compared to the monovalent counterpart. However, in cellulo, the nano-eGFP-E3 construct showed its superiority over the tandem format in terms of specificity with a highest and better ratio signal-to-noise. All together, the E3 peptide provides a universal suitable tool for the construction of dimeric biomolecules, in particular antibody fragments with improved functional affinity.
(Copyright © 2021 Elsevier B.V. All rights reserved.)
