Abstract
Antibody-Oligonucleotide Conjugates (AOCs) represent an emerging class of functionalized antibodies that have already been used in a wide variety of applications. While the impact of dye and drug conjugation on antibodies’ ability to bind their target has been extensively studied, little is known about the effect caused by the conjugation of hydrophilic and charged payloads such as oligonucleotides on the functions of an antibody. Previous observations of non-specific interactions of nucleic acids with untargeted cells prompted us to further investigate their impact on AOC binding abilities and cell selectivity. We synthesized a series of single- and double-stranded AOCs, as well as a human serum albumin-oligonucleotide conjugate, and studied their interactions with both targeted and non-targeted living cells using a time-resolved analysis of ligand binding assay. Our results indicate that conjugation of single strand oligonucleotides to proteins induce consistent non-specific interactions with cell surfaces while double strand oligonucleotides have little or no effect, depending on the preparation method.
Highlights
Antibody-Oligonucleotide Conjugates (AOCs) represent an emerging class of functionalized antibodies that have already been used in a wide variety of applications
As AOCs are developing into powerful tools in various applications[1], investigations to get a better understanding of their interactions with cell surfaces appear to be stimulating a renewed interest[30]
Our previous[2] and present results show that ONs are a particular payload that may display weak but consistent interactions with cell surfaces, which can impact the binding properties of antibodies upon conjugation
Summary
Antibody-Oligonucleotide Conjugates (AOCs) represent an emerging class of functionalized antibodies that have already been used in a wide variety of applications. Combining the specific binding ability of antibodies with the vast structural and functional properties of oligonucleotides (ONs), these conjugates have found a wide variety of applications as imaging, detection and therapeutic a gents[1] All of these functions primarily require the discrimination of the targeted cell type via specific binding of the AOC to its protein target. We observed that while our DNA-linked ADC (based on the anti-HER2 monoclonal antibody trastuzumab) showed a similar toxicity profile on HER2+ cells to classical covalent conjugates, it showed low but unexpected toxicity on the control HER2− cell line We hypothesized that this toxicity was the result of a non-specific interaction of the conjugate with HER2− cells induced by the ON linker. Despite these early warnings, the effect of ON conjugation on antibody selectivity remains understudied
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