Protein Rebinding to a Surface‐Confined Imprint

Dechtrirat, Decha; Jetzschmann, Katharina J.; Stöcklein, Walter; Scheller, Frieder W.; Gajovic‐Eichelmann, Nenad

doi:10.1002/adfm.201201328

Cited by 120 publications

(109 citation statements)

References 49 publications

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“…[33] Therefore, we were interested to explore the use of MIPs, that are expected to give a broader specificity than antibodies, for the quantitation of HSA in urine samples of patients showing microalbuminuria. While several electropolymerizable monomers emerged for protein imprinting applications [12] for preparing the HSA-imprinted MIP we used scopoletin as monomer, which has been introduced by Gajovich-Eichelmann [34] for the electrosynthesis of protein-MIPs and proved to enable the recognition of several protein targets. [34][35][36][37][38] By electropolymerization scopoletin forms an insulating polymer film, the thickness of which can be tuned to match the characteristic dimensions of the protein.…”

Section: Introductionmentioning

confidence: 99%

“…While several electropolymerizable monomers emerged for protein imprinting applications [12] for preparing the HSA-imprinted MIP we used scopoletin as monomer, which has been introduced by Gajovich-Eichelmann [34] for the electrosynthesis of protein-MIPs and proved to enable the recognition of several protein targets. [34][35][36][37][38] By electropolymerization scopoletin forms an insulating polymer film, the thickness of which can be tuned to match the characteristic dimensions of the protein. The protein binding to the MIP was detected by measuring the oxidative current of a redox probe on the underlying electrode, i.e., the protein binding hinders the permeability of the redox probe through the MIP nanofilm.…”

Section: Introductionmentioning

confidence: 99%

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Electrosynthesized molecularly imprinted polyscopoletin nanofilms for human serum albumin detection

Stojanović

Erdőssy

Keltai

et al. 2017

Analytica Chimica Acta

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Molecularly imprinted polymers (MIPs) rendered selective solely by the imprinting with protein templates lacking of distinctive properties to facilitate strong target-MIP interaction are likely to exhibit medium to low template binding affinities. While this prohibits the use of such MIPs for applications requiring the assessment of very low template concentrations, their implementation for the quantification of high-abundance proteins seems to have a clear niche in the analytical practice. We investigated this opportunity by developing a polyscopoletin-based MIP nanofilm for the electrochemical determination of elevated human serum albumin (HSA) in urine. As reference for low abundance protein ferritin-MIPs were 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 2 also prepared by the same procedure. Under optimal conditions, the imprinted sensors gave a linear response to HSA in the concentration range of 20-100 mg/dm 3 , and to ferritin in the range of 120-360 mg/dm 3 . While as expected the obtained limit of detection was not sufficient to determine endogenous ferritin in plasma, the HSA-sensor was successfully employed to analyse urine samples of patients with albuminuria. The results suggest that MIP-based sensors may be applicable for quantifying high abundance proteins in a clinical setting.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Electrosynthesized molecularly imprinted polyscopoletin nanofilms for human serum albumin detection

Stojanović

Erdőssy

Keltai

et al. 2017

Analytica Chimica Acta

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“…Scopoletin was deposited from aqueous solution forming an electrically non-conducting and hydrophilic film with a thickness ranging from 3 to 20 nm, depending on the monomer concentration and the polymerization method (Dechtrirat et al, 2012) (Bosserdt et al, 2015).…”

Section: Preparation Of Mips and Nipsmentioning

confidence: 99%

Electrosynthesized MIPs for transferrin: Plastibodies or nano-filters?

Zhang

Yarman

Erdőssy

et al. 2018

Biosensors and Bioelectronics

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Molecularly imprinted polymer (MIP) nanofilms for transferrin (Trf) have been synthesized on gold surfaces by electro-polymerizing the functional monomer scopoletin in the presence of the protein target or around pre-adsorbed Trf. As determined by atomic force microscopy (AFM) the film thickness was comparable with the molecular dimension of the target. The target (re)binding properties of the electro-synthesized MIP films was evaluated by cyclic voltammetry (CV) and square wave voltammetry (SWV) through the target-binding induced permeability changes of the MIP nanofilms to the ferricyanide redox marker, as well as by surface plasmon resonance (SPR) and surface enhanced infrared absorption spectroscopy (SEIRAS) of the immobilized protein molecules. For Trf a linear concentration dependence in the lower micromolar range and an imprinting factor of ~5 was obtained by 2 SWV and SPR. Furthermore, non-target proteins including the iron-free apo-Trf were discriminated by pronounced size and shape specificity. Whilst it is generally assumed that the rebinding of the target or of cross-reacting proteins exclusively takes place at the polymer here we considered also the interaction of the protein molecules with the underlying gold transducers. We demonstrate by SWV that adsorption of proteins suppresses the signal of the redox marker even at the bare gold surface and by SEIRAS that the treatment of the MIP with proteinase K or NaOH only partially removes the target protein. Therefore, we conclude that when interpreting binding of proteins to directly MIP-covered gold electrodes the interactions between the protein and the gold surface should also be considered.

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“…However, for biomacromolecular targets such as proteins, this still remains challenging due to their large size, high surface complexity, and conformational flexibility. Better accessibility of large target molecules can be achieved by generating the binding sites directly at the surface [31]. Various so-called surface-imprinting techniques were developed in the past few years employing in particular (photo)chemical polymerization, electrochemical synthesis, self-polymerization of dopamine by ambient oxygen, as well as enzyme-initiated polymerization (see Scheme 1).…”

Section: Preparation Of Surface Imprinted Mipsmentioning

confidence: 99%

“…In fact, the choice of regeneration conditions is a trade-off between complete removal of the target and preservation of the integrity of the binding sites. Methods applied include the application of chaotropic agents, extraction by organic solvents, the use of highly acidic or basic solutions and/or surfactants such as sodium dodecylsulfate (SDS) or Tween 20, sometimes at elevated temperatures [7], and electroelution [10,13,31].…”

Section: Template Removal By Enzymesmentioning

confidence: 99%

Enzymes as Tools in MIP-Sensors

et al. 2017

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Abstract:Molecularly imprinted polymers (MIPs) have the potential to complement antibodies in bioanalysis, are more stable under harsh conditions, and are potentially cheaper to produce. However, the affinity and especially the selectivity of MIPs are in general lower than those of their biological pendants. Enzymes are useful tools for the preparation of MIPs for both low and high-molecular weight targets: As a green alternative to the well-established methods of chemical polymerization, enzyme-initiated polymerization has been introduced and the removal of protein templates by proteases has been successfully applied. Furthermore, MIPs have been coupled with enzymes in order to enhance the analytical performance of biomimetic sensors: Enzymes have been used in MIP-sensors as "tracers" for the generation and amplification of the measuring signal. In addition, enzymatic pretreatment of an analyte can extend the analyte spectrum and eliminate interferences.

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Protein Rebinding to a Surface‐Confined Imprint

Abstract: 100% coverage 60% 0.05 mM MFI / a.u. c /mM 0.05 mM

Cited by 120 publications

References 49 publications

Electrosynthesized molecularly imprinted polyscopoletin nanofilms for human serum albumin detection

Electrosynthesized molecularly imprinted polyscopoletin nanofilms for human serum albumin detection

Electrosynthesized MIPs for transferrin: Plastibodies or nano-filters?

Enzymes as Tools in MIP-Sensors

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