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In article number 1807332, Zeynep Altintas and co‐workers combine computational and experimental studies to develop high affinity artificial protein binders by targeting a cancer biomarker. The artificial receptors show excellent affinity toward the protein marker, even superior to that of state‐of‐the‐art antibodies. [ABSTRACT FROM AUTHOR]

Widely used diagnostic tools make use of antibodies recognizing targeted molecules, but additional techniques are required in order to alleviate the disadvantages of antibodies. Herein, molecular dynamic calculations are performed for the design of high affinity artificial protein binding surfaces for the recognition of neuron specific enolase (NSE), a known cancer biomarker. Computational simulations are employed to identify particularly stabile secondary structure elements. These epitopes are used for the subsequent molecular imprinting, where surface imprinting approach is applied. The molecular imprints generated with the calculated epitopes of greater stability (Cys‐Ep1) show better binding properties than those of lower stability (Cys‐Ep5). The average binding strength of imprints created with stabile epitopes is found to be around twofold and fourfold higher for the NSE derived peptide and NSE protein, respectively. The recognition of NSE is investigated in a wide concentration range, where high sensitivity (limit of detection (LOD) = 0.5 ng mL−1) and affinity (dissociation constant (Kd) = 5.3 × 10−11m) are achieved using Cys‐Ep1 imprints reflecting the stable structure of the template molecules. This integrated approach employing stability calculations for the identification of stabile epitopes is expected to have a major impact on the future development of high affinity protein capturing binders. Combination of computational and experimental studies to develop high affinity artificial protein binders by targeting a cancer biomarker. Highly stable and least stable epitopes of neuron specific enolase are determined by molecular dynamic calculations, and they are used as templates for molecular imprinting. Molecular imprints created with stable epitopes are used as high affinity protein binders, while unsuitable epitope candidates are eliminated. [ABSTRACT FROM AUTHOR]

Abstract This research aims to engineer molecularly imprinted polymer (MIP)-based synthetic receptors for the molecular recognition of neuron specific enolase (NSE) biomarker. The synthetic peptide derived from the NSE was synthesized along with its cysteine and histidine modified versions. The modified peptides were utilized as templates for molecular imprinting, which was achieved by combination of epitope- and electrochemical surface imprinting strategy. The subsequently generated imprinted cavities were used for the detection of the NSE derived peptide and NSE. The imprints created with cysteine (CME) and histidine modified epitopes (HME) could detect the peptide in a concentration range of 2–128 µM and 15.6 nM to 128 µM, respectively. The recognition of NSE was achieved by the same imprints in a linear range of 1–64 ng mL−1 (CME) and 0.25–64 ng mL−1 (HME), respectively. The target molecules bound to the control polymer very weakly, confirming the high selectivity of the MIP cavities. Selectivity studies resulted in imprinting factors of 8.8 and 11 for the CME and HME imprints, respectively. The affinity analyses provided dissociation constants of 2.3 × 10−10 M and 3 × 10–11 M for NSE recognition using the corresponding epitope imprints. Cross-reactivity studies with non-specific molecules proved high specificity of the artificial receptors for the targets. Highlights • First epitope-mediated MIPs synthesized for neuron specific enolase biomarker. • Two different epitope imprinting approaches using cysteine- and histidine-modified peptides. • Highly specific and sensitive electrochemical sensors were manufactured with epitope imprints. • CV, SWV and AFM were employed to reveal template-MIP interactions. • This universal and promising strategy can be employed for a wide range protein markers. [ABSTRACT FROM AUTHOR]