Combinatorial screening of polymer nanoparticles for their ability to recognize epitopes of AAV‐neutralizing antibodies

Abstract

A library of 17 nanoparticles made of acrylate and methacrylate copolymers is prepared, characterized, and screened against six epitopes of adeno‐associated viruses (AAV)‐neutralizing antibodies to assess their affinity and specificity. Peptide epitopes are immobilized onto the surface of glass beads, packed in filtration microplates, and incubated with fluorescein‐labelled nanoparticles. Following intense washing, the affinity of nanoparticles to immobilized epitopes is assessed by measuring the fluorescence of captured nanoparticles. The results show that polar monomers, acrylic acid in particular, have a positive impact on polymer affinity towards all peptides used in this study. The presence of hydrophobic monomers, on other hand, has a negative impact on polymer binding. The composition of peptides used in this study has no noticeable impact on the affinity of synthesized nanoparticles. The affinity of nanoparticles with the highest affinity to peptide targets does not exceed millimolar level. Overall, it is found that the synthesized library showed modest affinity but lacked specificity, which should be further “tuned,” for example, by using molecular imprinting to achieve an acceptable level of affinity and specificity for practical application.

Advances in Electrochemical Nano-Biosensors for Biomedical and Environmental Applications: From Current Work to Future Perspectives

“…Numerous researches addressed the use of MIPs for accurate viral diagnosis in real-time [ 126 ]. MIPs fabricated by Piletska et al were applied for virus identification and the screening of different epitopes of Adeno-associated viruses (AAV) immobilized on glass beads using polymeric nanoparticles of acrylate and methacrylate [ 127 ]. On the other hand, a biosensor was fabricated for the viral hexon protein (the most accessible and abundant surface protein of the human adenovirus type 5 (hAdV5) icosahedral capsid).…”

The Amount of Cross-Linker Influences Affinity and Selectivity of NanoMIPs Prepared by Solid-Phase Polymerization Synthesis

“…At the end of the polymerization process, the non-covalent interaction between nanoMIPs and grafted template molecules is strong enough to allow any residual monomers, polymerization by-products, and low-affinity polymers to be easily removed by washing the glass beads with a weak solvent—typically cold water—whereas high-affinity nanoMIPs are subsequently eluted by washing with a stronger solvent able to break the non-covalent molecular interactions. When performed in water, the SPPS approach has shown its validity for very different types of polar templates, such as small organic molecules [ 10 , 11 , 12 , 13 , 14 ], peptides and proteins [ 15 , 16 , 17 , 18 , 19 ], polysaccharides [ 20 , 21 , 22 ], nucleic acids [ 23 ], viruses [ 24 , 25 ], and whole cells [ 26 ].…”