Modulation of EGFR Activity by Molecularly Imprinted Polymer Nanoparticles Targeting Intracellular Epitopes

Abstract

In recent years, molecularly imprinted polymer nanoparticles (nanoMIPs) have proven to be an attractive alternative to antibodies in diagnostic and therapeutic applications. However, several key questions remain: how suitable are intracellular epitopes as targets for nanoMIP binding? And to what extent can protein function be modulated via targeting specific epitopes? To investigate this, three extracellular and three intracellular epitopes of epidermal growth factor receptor (EGFR) were used as templates for the synthesis of nanoMIPs which were then used to treat cancer cells with different expression levels of EGFR. It was observed that nanoMIPs imprinted with epitopes from the intracellular kinase domain and the extracellular ligand binding domain of EGFR caused cells to form large foci of EGFR sequestered away from the cell surface, caused a reduction in autophosphorylation, and demonstrated effects on cell viability. Collectively, this suggests that intracellular domain-targeting nanoMIPs can be a potential new tool for cancer therapy.

Molecularly imprinted nanoparticles reveal regulatory scaffolding features in Pyk2 tyrosine kinase

“…, low nanomolar dissociation constants) and distinguish closely related amino acids such as leucine/isoleucine, 18 aspartate/glutamate, 20 and lysine/arginine. 21 The capacity for exquisite specificity enables MINPs to inhibit specific post-translational modifications 8,22 or probe the functional role of a specific peptide sequence. 19 Hence, we investigated the mechanisms regulating Pyk2 activation by leveraging the distinctive features of MINPs to selectively target and probe specific Pyk2 regulatory features (Fig.…”

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 ].…”

Molecularly imprinted nanoparticles reveal regulatory scaffolding features in Pyk2 tyrosine kinase

“…The resultant “plastic antibodies” can bind peptides with high affinity (i.e., low nanomolar dissociation constants) and distinguish closely related amino acids such as leucine/isoleucine 18 , aspartate/glutamate 20 , and lysine/arginine 21 . The capacity for exquisite specificity enables MINPs to inhibit specific post-translational modifications 8,22 or probe the functional role of a specific peptide sequence 19 . Hence, we investigated the mechanisms regulating Pyk2 activation by leveraging the distinctive features of MINPs to selectively target and probe specific Pyk2 regulatory features (Fig.…”