Conjugation of Antibodies and siRNA Duplexes to Polymer Nanoparticles via Maleimide–Thiol Chemistry
Date
2024-11-18
Journal Title
Journal ISSN
Volume Title
Publisher
ACS Omega
Abstract
Polymeric nanoparticles (NPs) have shown great promise as highly modifiable platforms that can be applied across many different disease states. They are advantageous because they can encapsulate a range of hydrophobic and hydrophilic cargoes while having customizable surface properties. Depending on the desired biointerfacing capabilities, the surface of polymeric NPs can be modified with moieties, such as antibodies, peptides, nucleic acids, and more. The work presented here is intended to provide mechanistic insight into how different parameters influence the loading of antibodies, small interfering ribonucleic acids (siRNAs), or both on the surface of poly(lactic-co-glycolic acid) (PLGA) NPs via maleimide–thiol chemistry. Some of the conjugation parameters investigated include the buffer concentration, maleimide to protein ratio, and the addition of an excipient such as Tween-20. Through variation in the concentration of FZD7 antibodies added to the reaction mixture, we established tunable conjugation and found the upper limit of their loading density under the conditions tested. We also confirmed antibody conjugation through two different mechanisms: via a thiol-modified antibody or a thiol-modified poly(ethylene glycol) (PEG) linker. Conjugation of thiolated siRNA duplexes targeting β-catenin was also investigated through variations in both Tween-20 concentration and CaCl2 buffer concentration. Finally, the coconjugation of both antibodies and siRNA duplexes was explored. Overall, this work outlines a basis for tunable biomolecule loading on polymer NPs using maleimide–thiol chemistry and reveals the incredible versatility of polymer NP platforms.
Description
This article was originally published in ACS Omega. The version of record is available at: https://doi.org/10.1021/acsomega.4c07025.
© 2024 The Authors. Published by American Chemical Society. This publication is licensed under CC-BY-NC-ND 4.0 (https://creativecommons.org/licenses/by-nc-nd/4.0/).
Keywords
biopolymers, genetics, immunology, molecular properties, peptides and proteins
Citation
Hoover, Elise C., Chitran Roy Chowdhury, Olivia M. Ruggiero, and Emily S. Day. “Conjugation of Antibodies and siRNA Duplexes to Polymer Nanoparticles via Maleimide–Thiol Chemistry.” ACS Omega, November 18, 2024, acsomega.4c07025. https://doi.org/10.1021/acsomega.4c07025.