Protein‐Templated Reactions Using DNA‐Antibody Conjugates

Abstract

DNA-templated chemical reactions have found wide applications in drug discovery, programmed multistep synthesis, nucleic acid detection and targeted drug delivery. The control of these reactions has, however, been limited to nucleic acid hybridization as a means to direct the proximity between reactants.In this work we introduce a system capable of translating protein-protein binding events into a DNAtemplated reaction which leads to the covalent formation of a product. We achieve protein-templated reactions by employing two DNA-antibody conjugates that are both able to recognize the same target protein and to colocalize a pair of reactant DNA strands able to undergo a click reaction. We engineered two individual systems each responsive to human serum albumin (HSA) and human IgG and we demonstrated that, while no reaction occurs in the absence of proteins, both protein-templated reactions can occur simultaneously in the same solution without any inter-system crosstalk.

Synthetic Antigen-Conjugated DNA Systems for Antibody Detection and Characterization

“…The versatility of our approach leads not only to trigger a second reaction (phosphoramidate ligation reaction) using a different recognition element/antibody couple but also to achieve an orthogonal control of two different templated reactions in the same solution with two different antibodies (Figure B). More recently, in a follow-up work in collaboration with the Gothelf group, we have extended the same approach to translate protein–protein binding events into DNA-templated reactions . Antibody-controlled templated reactions can be also useful for sensing applications in case the reaction leads to a measurable output (for example optical), but many other possible applications can be envisioned.…”

A Proximity‐Induced Fluorogenic Reaction Triggered by Antibody–Antigen Interactions with Adjacent Epitopes

“…[1] In the presence of templates to induce this proximity, local concentration of the substrates is increased by specific and strong interactions, thus promoting the desired reaction even in multimolecular systems. A class of well-known examples is the DNA-templated chemical reaction, [2][3][4][5][6][7][8][9] in which reactive functional groups linked to DNA are brought into proximity by complementary hybridization via duplex formation. [4,5] Various chemical reactions have been demonstrated for nucleic acid detection, [6,7] drug release, [8] and small-molecule drug discovery.…”

“…A class of well-known examples is the DNA-templated chemical reaction, [2][3][4][5][6][7][8][9] in which reactive functional groups linked to DNA are brought into proximity by complementary hybridization via duplex formation. [4,5] Various chemical reactions have been demonstrated for nucleic acid detection, [6,7] drug release, [8] and small-molecule drug discovery. [9] In some other approaches, proximity is gained by using interactions of oligonucleotidepeptide nucleic acid, [10,11] peptide-peptide [12] and proteinpeptide.…”

“…[13] Polyclonal antibody-antigen interactions have also been combined to bring DNA in close proximity to induce hybridization at lower concentrations. [4] Antibody-antigen interactions are specific, strong, and potentially suitable for proximity-induced chemical reactions. We recently reported that a biparatopic antibody can be generated on an antigen with the assistance of peptideprotein complex formation.…”

Editorial for the Special Issue ‘Synthetic Biology and Biomimicry'

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Ricci, Gothelf et al present DNA-antibody conjugates that use biomolecular recognition to perform protein template reactions. [7] Zaburdaev, Göpfrich et al approach the challenge of division of synthetic cells and investigate segregation of DNA within cellular mimics. [8] Andersen et al report on the cotranscriptional assembly of RNA into nanostructures with stabilizing geometries.

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