A Platform for Site‐Specific DNA‐Antibody Bioconjugation by Using Benzoylacrylic‐Labelled Oligonucleotides
Open Access
- 23 September 2021
- journal article
- research article
- Published by Wiley in Angewandte Chemie
- Vol. 60 (49), 25905-25913
- https://doi.org/10.1002/anie.202109713
Abstract
Many bioconjugation strategies for DNA oligonucleotides and antibodies suffer limitations, such as site-specificity, stoichiometry and hydrolytic instability of the conjugates, which makes them unsuitable for biological applications. Here, we report a new platform for the preparation of DNA-antibody bioconjugates with a simple benzoylacrylic acid pentafluorophenyl ester reagent. Benzoylacrylic-labelled oligonucleotides prepared with this reagent can be site-specifically conjugated to a range of proteins and antibodies through accessible cysteine residues. The homogeneity of the prepared DNA-antibody bioconjugates was confirmed by a new LC–MS protocol and the bioconjugate probes were used in fluorescence or super-resolution microscopy cell imaging experiments. This work demonstrates the versatility and robustness of our bioconjugation protocol that gives site-specific, well-defined and plasma-stable DNA-antibody bioconjugates for biological applications.Keywords
Funding Information
- Biotechnology and Biological Sciences Research Council (iCASE)
- H2020 European Research Council (676832)
- Royal Society (RP150066, URF\R\180019)
- Fundação para a Ciência e a Tecnologia (CEECIND/00453/2018)
This publication has 63 references indexed in Scilit:
- Orthogonal Protein Decoration of DNA NanostructuresSmall, 2011
- Tunable Degradation of Maleimide–Thiol Adducts in Reducing EnvironmentsBioconjugate Chemistry, 2011
- Functionalization of DNA nanostructures with proteinsChemical Society Reviews, 2011
- Comparison of the C2A Domain of Synaptotagmin-I and Annexin-V As Probes for Detecting Cell DeathBioconjugate Chemistry, 2010
- Semisynthetic DNA–Protein Conjugates for Biosensing and NanofabricationAngewandte Chemie, 2010
- Halbsynthetische DNA‐Protein‐Konjugate für Biosensorik und NanofabrikationAngewandte Chemie, 2010
- Expression, purification and use of recombinant annexin V for the detection of apoptotic cellsNature Protocols, 2009
- Comparison of LC and LC/MS methods for quantifying N-glycosylation in recombinant IgGsJournal of the American Society for Mass Spectrometry, 2008
- Imaging individual mRNA molecules using multiple singly labeled probesNature Methods, 2008
- Site-specific conjugation of a cytotoxic drug to an antibody improves the therapeutic indexNature Biotechnology, 2008