Epitope recognition of peptide-imprinted polymers for Regenerating protein 1 (REG1)

Abstract

Molecularly imprinted polymers (MIPs) were developed to replace natural antibodies with a cost-effective and durable synthetic material. Molecular imprinting of proteins conventionally utilizes the whole protein as the template, which is complex (as many different epitopes may be imprinted) and expensive. In this work, seven peptides (13-18 amino acids) were synthesized and used as templates for the imprinting and recognition of Regenerating Protein 1 (REG1). REG1 is involved in the proliferation and differentiation of diverse cell types, and was recently described as a urinary biomarker for pancreatic ductal adenocarcinoma (PDAC). Peptide-imprinted poly(ethylene-co-vinyl alcohol)s (PIPs), containing four different mole fractions of ethylene were cast on screen-printed electrodes to find the optimum composition for both the sensing and the extraction of REG1 in an E coli culture medium. Peptides with fewer than 16 amino acids and two or three aromatic and hydrophobic groups have a higher affinity for MIPs of poly(ethylene-co-vinyl alcohol) (EVAL) with 27 mole% of ethylene, while those with four aromatic and hydrophobic groups have a higher affinity for MIPs with EVALs that contain 32 mole% of ethylene. The peptide / EVAL combination that maximized both imprinting effectiveness and response to REG1B was the sequence NEDRETWVDADLY imprinted into 32 mol% EVAL. This EVAL composition and template peptide were then modified by incorporation of magnetic nanoparticles, thus extending applications for PIPs to include extraction of REG1 protein from E coli culture medium.

Immunotherapy of Hepatocellular Carcinoma with Magnetic PD-1 Peptide-Imprinted Polymer Nanocomposite and Natural Killer Cells

“…Thus, inexpensive proteins such as albumin [20], lysozyme [21], or hemoglobin were employed in model systems. Recently, useful peptide epitopes of proteins were found via molecular simulation [22] or empirical methods [23], and used as templates in molecular imprinting. Peptide-imprinted polymers were characterized and found to achieve both high capacity and selectivity [23].…”

“…Recently, useful peptide epitopes of proteins were found via molecular simulation [22] or empirical methods [23], and used as templates in molecular imprinting. Peptide-imprinted polymers were characterized and found to achieve both high capacity and selectivity [23]. Peptides that contain six to 14 amino acids are typically selected as epitope templates for imprinting [22].…”

Immunotherapy of Hepatocellular Carcinoma with Magnetic PD-1 Peptide-Imprinted Polymer Nanocomposite and Natural Killer Cells

“…Recently, peptide epitopes of proteins have been found by molecular simulation 21 or empirical methods, 22 and used as templates in molecular imprinting. Peptide-imprinted polymers have been characterized and found to achieve both high capacity and selectivity.…”

“…Peptide-imprinted polymers have been characterized and found to achieve both high capacity and selectivity. 22 Peptides that contain six to 14 amino acids are typically selected as epitope templates for imprinting. 21 However, there are no general rules for peptide selection that involve the various functions of proteins, including catalysis, cell signaling and ligand binding, and protein structure.…”

Cellular reprogramming with multigene activation by the delivery of CRISPR/dCas9 ribonucleoproteins via magnetic peptide-imprinted chitosan nanoparticles

“…Magnetic molecularly imprinted polymers (MMIPs) are magnetic nanoparticles (MNPs) that are coated with tailor-made recognition polymers for biosensing [ 12 ], bioseparation [ 13 , 14 ] and delivery [ 13 , 15 , 16 ]. In our previous work, seven peptides (13–18 amino acids) were synthesized and used as templates for the imprinting and recognition of Regenerating Protein 1 [ 14 ]. Recently, one peptide of programmed cell death protein 1 was used as the template for molecular imprinting to form magnetic peptide-imprinted poly(ethylene- co -vinyl alcohol) composite nanoparticles [ 16 ].…”