TY - JOUR A1 - Ozcelikay, Goksu A1 - Kurbanoglu, Sevinc A1 - Yarman, Aysu A1 - Scheller, Frieder W. A1 - Ozkan, Sibel A. T1 - Au-Pt nanoparticles based molecularly imprinted nanosensor for electrochemical detection of the lipopeptide antibiotic drug Daptomycin JF - Sensors and actuators : B, Chemical N2 - In this work, a novel electrochemical molecularly imprinted polymer (MIP) sensor for the detection of the lipopeptide antibiotic Daptomycin (DAP) is presented which integrates gold decorated platinum nanoparticles (Au-Pt NPs) into the nanocomposite film. The sensor was prepared by electropolymerization of o-phenylenediamine (o-PD) in the presence of DAP using cyclic voltammetry. Cyclic voltammetry and differential pulse voltammetry were applied to follow the changes in the MIP-layer related to rebinding and removal of the target DAP by using the redox marker [Fe(CN)(6)](3-/4-). Under optimized operational conditions, the MIP/Au-Pt NPs/ GCE nanosensor exhibits a linear response in the range of 1-20 pM towards DAP. The limit of detection and limit of quantification were determined to be 0.161pM +/- 0.012 and 0.489pM +/- 0.012, respectively. The sensitivity towards the antibiotics Vancomycin and Erythromycin and the amino acids glycine and tryptophan was below 7 percent as compared with DAP. Moreover, the nanosensor was also successfully used for the detection of DAP in deproteinated human serum samples. KW - molecularly imprinted polymer KW - Daptomycin KW - platinum nanoparticles KW - gold KW - nanoparticles KW - modified electrodes Y1 - 2020 U6 - https://doi.org/10.1016/j.snb.2020.128285 SN - 0925-4005 VL - 320 PB - Elsevier Science CY - Amsterdam ER - TY - GEN A1 - Yarman, Aysu A1 - Scheller, Frieder W. T1 - How reliable is the electrochemical readout of MIP-sensors? T2 - Postprints der Universität Potsdam : Mathematisch Naturwissenschaftliche Reihe N2 - Electrochemical methods offer the simple characterization of the synthesis of molecularly imprinted polymers (MIPs) and the readouts of target binding. The binding of electroinactive analytes can be detected indirectly by their modulating effect on the diffusional permeability of a redox marker through thin MIP films. However, this process generates an overall signal, which may include nonspecific interactions with the nonimprinted surface and adsorption at the electrode surface in addition to (specific) binding to the cavities. Redox-active low-molecular-weight targets and metalloproteins enable a more specific direct quantification of their binding to MIPs by measuring the faradaic current. The in situ characterization of enzymes, MIP-based mimics of redox enzymes or enzyme-labeled targets, is based on the indication of an electroactive product. This approach allows the determination of both the activity of the bio(mimetic) catalyst and of the substrate concentration. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 960 KW - molecularly imprinted polymers KW - electropolymerization KW - direct electron transfer KW - catalysis KW - redox marker KW - gate effect Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-471608 SN - 1866-8372 IS - 960 ER - TY - JOUR A1 - Yarman, Aysu A1 - Kurbanoğlu, Sevinç A1 - Zebger, Ingo A1 - Scheller, Frieder W. T1 - Simple and robust BT - the claims of protein sensing by molecularly imprinted polymers JF - Sensors and actuators : B, Chemical : an international journal devoted to research and development of chemical transducers N2 - A spectrum of 7562 publications on Molecularly Imprinted Polymers (MIPs) has been presented in literature within the last ten years (Scopus, September 7, 2020). Around 10 % of the papers published on MIPs describe the recognition of proteins. The straightforward synthesis of MIPs is a significant advantage as compared with the preparation of enzymes or antibodies. MIPs have been synthesized from only one up to six functional monomers while proteins are made up of 20 natural amino acids. Furthermore, they can be synthesized against structures of low immunogenicity and allow multi-analyte measurements via multi-target synthesis. Electrochemical methods allow simple polymer synthesis, removal of the template and readout. Among the different sensor configurations electrochemical MIP-sensors provide the broadest spectrum of protein analytes. The sensitivity of MIP-sensors is sufficiently high for biomarkers in the sub-nanomolar region, nevertheless the cross-reactivity of highly abundant proteins in human serum is still a challenge. MIPs for proteins offer innovative tools not only for clinical and environmental analysis, but also for bioimaging, therapy and protein engineering. KW - Molecularly imprinted polymer KW - Plastibodies KW - Functional scaffolds KW - Biomimetic sensors KW - Proteins Y1 - 2021 U6 - https://doi.org/10.1016/j.snb.2020.129369 SN - 0925-4005 SN - 1873-3077 VL - 330 PB - Elsevier Science CY - Amsterdam [u.a.] ER - TY - JOUR A1 - Caserta, Giorgio A1 - Zhang, Xiaorong A1 - Yarman, Aysu A1 - Supala, Eszter A1 - Wollenberger, Ulla A1 - Gyurcsányi, Róbert E. A1 - Zebger, Ingo A1 - Scheller, Frieder W. T1 - Insights in electrosynthesis, target binding, and stability of peptide-imprinted polymer nanofilms JF - Electrochimica acta : the journal of the International Society of Electrochemistry (ISE) N2 - Molecularly imprinted polymer (MIP) nanofilms have been successfully implemented for the recognition of different target molecules: however, the underlying mechanistic details remained vague. This paper provides new insights in the preparation and binding mechanism of electrosynthesized peptide-imprinted polymer nanofilms for selective recognition of the terminal pentapeptides of the beta-chains of human adult hemoglobin, HbA, and its glycated form HbA1c. To differentiate between peptides differing solely in a glucose adduct MIP nanofilms were prepared by a two-step hierarchical electrosynthesis that involves first the chemisorption of a cysteinyl derivative of the pentapeptide followed by electropolymerization of scopoletin. This approach was compared with a random single-step electrosynthesis using scopo-letin/pentapeptide mixtures. Electrochemical monitoring of the peptide binding to the MIP nanofilms by means of redox probe gating revealed a superior affinity of the hierarchical approach with a Kd value of 64.6 nM towards the related target. Changes in the electrosynthesized non-imprinted polymer and MIP nanofilms during chemical, electrochemical template removal and rebinding were substantiated in situ by monitoring the characteristic bands of both target peptides and polymer with surface enhanced infrared absorption spectroscopy. This rational approach led to MIPs with excellent selectivity and provided key mechanistic insights with respect to electrosynthesis, rebinding and stability of the formed MIPs. KW - SEIRA spectroelectrochemistry KW - peptide imprinting KW - electrosynthesis KW - MIP KW - glycated peptide Y1 - 2021 U6 - https://doi.org/10.1016/j.electacta.2021.138236 SN - 0013-4686 SN - 1873-3859 VL - 381 PB - Elsevier CY - New York, NY [u.a.] ER - TY - JOUR A1 - Zhang, Xiaorong A1 - Caserta, Giorgio A1 - Yarman, Aysu A1 - Supala, Eszter A1 - Tadjoung Waffo, Armel Franklin A1 - Wollenberger, Ulla A1 - Gyurcsanyi, Robert E. A1 - Zebger, Ingo A1 - Scheller, Frieder W. T1 - "Out of Pocket" protein binding BT - a dilemma of epitope imprinted polymers revealed for human hemoglobin JF - Chemosensors N2 - The epitope imprinting approach applies exposed peptides as templates to synthesize Molecularly Imprinted Polymers (MIPs) for the recognition of the parent protein. While generally the template protein binding to such MIPs is considered to occur via the epitope-shaped cavities, unspecific interactions of the analyte with non-imprinted polymer as well as the detection method used may add to the complexity and interpretation of the target rebinding. To get new insights on the effects governing the rebinding of analytes, we electrosynthesized two epitope-imprinted polymers using the N-terminal pentapeptide VHLTP-amide of human hemoglobin (HbA) as the template. MIPs were prepared either by single-step electrosynthesis of scopoletin/pentapeptide mixtures or electropolymerization was performed after chemisorption of the cysteine extended VHLTP peptide. Rebinding of the target peptide and the parent HbA protein to the MIP nanofilms was quantified by square wave voltammetry using a redox probe gating, surface enhanced infrared absorption spectroscopy, and atomic force microscopy. While binding of the pentapeptide shows large influence of the amino acid sequence, all three methods revealed strong non-specific binding of HbA to both polyscopoletin-based MIPs with even higher affinities than the target peptides. KW - Molecularly Imprinted Polymers KW - epitope imprinting KW - non-specific KW - binding KW - redox gating KW - SEIRA spectroelectrochemistry Y1 - 2021 U6 - https://doi.org/10.3390/chemosensors9060128 SN - 2227-9040 VL - 9 IS - 6 PB - MDPI CY - Basel ER - TY - JOUR A1 - Bognár, Zsófia A1 - Supala, Eszter A1 - Yarman, Aysu A1 - Zhang, Xiaorong A1 - Bier, Frank Fabian A1 - Scheller, Frieder W. A1 - Gyurcsanyi, Róbert E. T1 - Peptide epitope-imprinted polymer microarrays for selective protein recognition BT - application for SARS-CoV-2 RBD protein JF - Chemical science / RSC, Royal Society of Chemistry N2 - We introduce a practically generic approach for the generation of epitope-imprinted polymer-based microarrays for protein recognition on surface plasmon resonance imaging (SPRi) chips. The SPRi platform allows the subsequent rapid screening of target binding kinetics in a multiplexed and label-free manner. The versatility of such microarrays, both as synthetic and screening platform, is demonstrated through developing highly affine molecularly imprinted polymers (MIPs) for the recognition of the receptor binding domain (RBD) of SARS-CoV-2 spike protein. A characteristic nonapeptide GFNCYFPLQ from the RBD and other control peptides were microspotted onto gold SPRi chips followed by the electrosynthesis of a polyscopoletin nanofilm to generate in one step MIP arrays. A single chip screening of essential synthesis parameters, including the surface density of the template peptide and its sequence led to MIPs with dissociation constants (K-D) in the lower nanomolar range for RBD, which exceeds the affinity of RBD for its natural target, angiotensin-convertase 2 enzyme. Remarkably, the same MIPs bound SARS-CoV-2 virus like particles with even higher affinity along with excellent discrimination of influenza A (H3N2) virus. While MIPs prepared with a truncated heptapeptide template GFNCYFP showed only a slightly decreased affinity for RBD, a single mismatch in the amino acid sequence of the template, i.e. the substitution of the central cysteine with a serine, fully suppressed the RBD binding. Y1 - 2021 U6 - https://doi.org/10.1039/d1sc04502d SN - 2041-6539 VL - 13 IS - 5 SP - 1263 EP - 1269 PB - Royal Society of Chemistry CY - Cambridge ER -