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A molecularly designed imprinting method was combined with a gravimetric nanosensor for the real-time detection Cu(II) ions in aqueous solutions without using expensive laboratory devices. Thus, 1:1 and 2:1 mol-ratio-dependent coordination modes between Cu(II), N-methacyloly-L histidine methyl ester (MAH) functional monomer complexes, and their four-fold and six-fold coordinations were calculated by means of density functional theory molecular modeling. Cu(II)-MIP1 and Cu(II)-MIP2 nanoparticles were synthesized in the size range of 80-100 nm and characterized by SEM, AFM and FTIR. Cu(II)-MIP nanoparticles were then conducted to a quartz crystal microbalance sensor for the real-time detection of Cu(II) ions in aqueous solutions. The effects of initial Cu(II) concentration, selectivity, and imprinting efficiency were investigated for the optimization of the nanosensor. Linearity of 99% was obtained in the Cu(II) ion linear concentration range of 0.15-1.57 µM with high sensitivity. The LOD was obtained as 40.7 nM for Cu(II)-MIP2 nanoparticles. The selectivity and the imprinting efficiency of the QCM nanosensor were obtained significantly in the presence of competitive ion samples (Co(II), Ni(II), Zn(II), and Fe(II)). The results are promising for sensing Cu(II) ions as environmental toxicants in water by combining molecularly designed ion-imprinted nanoparticles and a gravimetric sensor.

Considering the limitations of well-known traditional detection techniques, innovative research studies have focused on the development of new sensors to offer label-free, highly sensitive, real-time, low-cost, and rapid detection for biomolecular interactions. In this study, we demonstrate immunoglobulin G (IgG) detection in aqueous solutions by using real-time and label-free kinetic measurements of the Interferometric Reflectance Imaging Sensor (IRIS) system. By performing kinetic characterization experiments, the sensor's performance is comprehensively evaluated and a high correlation coefficient value (>0.94) is obtained in the IgG concentration range of 1-50 μg/mL with a low detection limit (0.25 μg/mL or 1.67 nM). Moreover, the highly sensitive imaging system ensures accurate quantification and reliable validation of recorded binding events, offering new perspectives in terms of direct biomarker detection for clinical applications., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The field of biosensing is in constant evolution, propelled by the need for sensitive, reliable platforms that provide consistent results, especially in the drug development industry, where small molecule characterization is of uttermost relevance. Kinetic characterization of small biochemicals is particularly challenging, and has required sensor developers to find solutions to compensate for the lack of sensitivity of their instruments. In this regard, surface chemistry plays a crucial role. The ligands need to be efficiently immobilized on the sensor surface, and probe distribution, maintenance of their native structure and efficient diffusion of the analyte to the surface need to be optimized. In order to enhance the signal generated by low molecular weight targets, surface plasmon resonance sensors utilize a high density of probes on the surface by employing a thick dextran matrix, resulting in a three-dimensional, multilayer distribution of molecules. Despite increasing the binding signal, this method can generate artifacts, due to the diffusion dependence of surface binding, affecting the accuracy of measured affinity constants. On the other hand, when working with planar surface chemistries, an incredibly high sensitivity is required for low molecular weight analytes, and furthermore the standard method for immobilizing single layers of molecules based on self-assembled monolayers (SAM) of epoxysilane has been demonstrated to promote protein denaturation, thus being far from ideal. Here, we will give a concise overview of the impact of tridimensional immobilization of ligands on label-free biosensors, mostly focusing on the effect of diffusion on binding affinity constants measurements. We will comment on how multilayering of probes is certainly useful in terms of increasing the sensitivity of the sensor, but can cause steric hindrance, mass transport and other diffusion effects. On the other hand, probe monolayers on epoxysilane chemistries do not undergo diffusion effect but rather other artifacts can occur due to probe distortion. Finally, a combination of tridimensional polymeric chemistry and probe monolayer is presented and reviewed, showing advantages and disadvantages over the other two approaches.

Molecularly imprinted polymer-based surface plasmon resonance sensor prepared using silver nanoparticles was designed for the selective recognition of Penicillin G (PEN-G) antibiotic from both aqueous solution and milk sample. PEN-G imprinted sensors (NpMIPs) SPR sensor was fabricated using poly (2-hydroxyethyl methacrylate-N-methacroyl-(L)-cysteine methyl ester)-silver nanoparticles-N-methacryloyl-L-phenylalanine methyl ester polymer by embedding silver nanoparticles (AgNPs) into the polymeric film structure. In addition, a non-imprinted (NpNIPs) SPR sensor was prepared by utilizing the same polymerization recipe without addition of the PEN-G template molecule to evaluate the imprinting effect. FTIR-ATR spectrophotometer, ellipsometer, contact angle measurements were used for the characterization of NpMIPs SPR sensors. The linear concentration range of 0.01-10 ng/mL PEN-G was studied for kinetic analyses. The augmenting effect of AgNPs used to increase the surface plasmon resonance signal response was examined using polymer-based PEN-G imprinted (MIPs) sensor without the addition of AgNPs. The antibiotic amount present in milk chosen as a real sample was measured by spiking PEN-G into the milk. According to the Scatchard, Langmuir, Freundlich and Langmuir-Freundlich adsorption models, the interaction mechanism was estimated to be compatible with the Langmuir model.

Microfluidic devices have led to novel biological advances through the improvement of micro systems that can mimic and measure. Microsystems easily handle sub-microliter volumes, obviously with guidance presumably through laminated fluid flows. Microfluidic systems have production methods that do not need expert engineering, away from a centralized laboratory, and can implement basic and point of care analysis, and this has attracted attention to their widespread dissemination and adaptation to specific biological issues. The general use of microfluidic tools in clinical settings can be seen in pregnancy tests and diabetic control, but recently microfluidic platforms have become a key novel technology for cancer diagnostics. Cancer is a heterogeneous group of diseases that needs a multimodal paradigm to diagnose, manage, and treat. Using advanced technologies can enable this, providing better diagnosis and treatment for cancer patients. Microfluidic tools have evolved as a promising tool in the field of cancer such as detection of a single cancer cell, liquid biopsy, drug screening modeling angiogenesis, and metastasis detection. This review summarizes the need for the low-abundant blood and serum cancer diagnosis with microfluidic tools and the progress that has been followed to develop integrated microfluidic platforms for this application in the last few years.

In this work, a surface plasmon resonance (SPR) based immunosensor was prepared by the immobilization of the amine-functionalized gold nanoparticles (N-AuNPs) on the sensing surface to sense immunoglobulin M (IgM) antibodies in the aqueous solution and artificial plasma. The characterization studies of SPR based immunosensor for IgM detection were performed with scanning electron microscope (SEM), contact angle measurements, and ellipsometry. Kinetic studies for the IgM immunosensor were carried out in the range of 1.0 to 200 ng/mL IgM concentrations in an aqueous solution. The total IgM analysis time including adsorption, desorption, and regeneration cycles was nearly 10 min for the prepared immunosensor. The limit of detection (LOD) and limit of quantification (LOQ) were found as 0.08 and 0.26 ng/mL, respectively. The reusability of the proposed immunosensor was tested with 6 consecutive adsorption-desorption, and regeneration cycles. Also, enzyme-linked immunosorbent assay (ELISA) method was utilized in the validation of the immunosensor.

Over the past few decades, a significant increase in multi-drug-resistant pathogenic microorganisms has been of great concern and directed the research subject to the challenges that the distribution of resistance genes represent. Globally, high levels of multi-drug resistance represent a significant health threat and there is a growing requirement of rapid, accurate, real-time detection which plays a key role in tracking of measures for the infections caused by these bacterial strains. It is also important to reduce transfer of resistance genes to new organisms. The, World Health Organization has informed that millions of deaths have been reported each year recently. To detect the resistant organisms traditional detection approaches face limitations, therefore, newly developed technologies are needed that are suitable to be used in large-scale applications. In the present study, the aim was to design a surface plasmon resonance (SPR) sensor with micro-contact imprinted sensor chips for the detection of Staphylococcus aureus . Whole cell imprinting was performed by N -methacryloyl-L-histidine methyl ester (MAH) under UV polymerization. Sensing experiments were done within a concentration range of 1.0 × 10 2 -2.0 × 10 5 CFU/mL. The recognition of S. aureus was accomplished by the involvement of microcontact imprinting and optical sensor technology with a detection limit of 1.5 × 10 3 CFU/mL. Selectivity of the generated sensor was evaluated through injections of competing bacterial strains. The responses for the different strains were compared to that of S. aureus. Besides, real experiments were performed with milk samples spiked with S. aureus and it was demonstrated that the prepared sensor platform was applicable for real samples.

In this study, composite polymeric cryogel cartridges were achieved by using Cd(II) imprinted poly(hydroxyethyl methacrylate N-methacryloly-(L)-cysteine methylester) beads and poly(hydroxyethyl methacrylate) cryogel cartridges with two different mole ratios of functional monomer. The N-methacryloly-(L)-cysteinemethylester was used as a functional monomer and Cd(II) 1:1 and 2:1, which were then notated as MIP1 and MIP2, respectively. Various characterization methods have confirmed the structural transformation on the MIP1 and MIP2 composite cryogel cartridges by scanning electron microscopy, Fourier-transform infrared spectroscopy-Attenuated Total Reflectance, and swelling tests. The maximum amount of Cd(II) adsorption with composite cryogel cartridges was determined by altering the Cd(II) initial concentration, temperature, and pH values. The maximum adsorption capacity of MIP1 and MIP2 composite cryogel cartridges obtained was 76.35 and 98.8 µmol/g of composite cryogels, respectively. The adsorption studies revealed that the MIP2 possessed a good adsorption performance for Cd(II). The obtained composite cryogel cartridges have a selective, reusable, and cost-friendly potential for the removal of Cd(II) from aqueous solutions, and are used many times without decreasing their adsorption capacities significantly. The Cd(II) removal rate of the MIP1 and MIP2 composite cryogel cartridges from synthetic wastewater samples was determined as 98.8%. The obtained cryogel cartridges' adsorption material exhibited a good directional removal performance for Cd(II) from wastewater samples.

Human immunoglobulin M (hIgM) antibodies are considered as hopeful tools for diseases therapy. Therefore, chromatography approaches are used to purify hIgM with a single step. In this study, we prepared a poly(hydroxyethyl methacrylate) based immunoaffinity p(HEMA-I) cryogel column by using cyanamide to immobilize antihuman immunoglobulin on the p(HEMA) cryogel for purification of hIgM in aqueous solution and artificial human plasma. The characterization of the p(HEMA) cryogel column was performed by using a scanning electron microscope (SEM), micro-computerized tomography (µ-CT), Fourier transform infrared spectroscopy (FTIR), swelling degree and macro-porosity. Further, the optimizations of various parameters were performed such as, pH, ionic strength, temperature and concentration of hIgM in aqueous solutions. In addition, the Langmuir adsorption model was supported by experimental results. Maximum adsorbed amount of hIgM corresponded to 11.1 mg/g at pH 5.75 [morpholino ethanesulfonic acid (MES buffer)]. Our results indicated that the p(HEMA-I) cryogel column can be reused at least 10 times without significant loss in adsorption capacity. As a natural source, artificial human plasma was selected for hIgM adsorption and the purity of hIgM was evaluated using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)., Competing Interests: There is no conflict of interest for authors.

There is a significant need for the development of the novel adsorbents in the field of protein purification. In this study, thin hemoglobin imprinted film (MIP) was fabricated onto the bacterial cellulose nanofibers' (BCNFs) by surface imprinting method using metal ion coordination interactions with N-methacryloyl-(L)-histidinemethylester (MAH) and copper ions. The hemoglobin surface imprinted bacterial cellulose nanofibers (MIP-BCNFs) was applied to selective recognition of hemoglobin and purification from hemolysate. The characterization of the MIP-BCNFs was carried out by the Fourier Transformed Infrared Spectroscopy (FT-IR), Scanning Electron Microscopy (SEM), micro-Computerized Tomography (μCT), atomic force microscopy (AFM) and surface area measurements. The adsorption experiments of hemoglobin onto the MIP-BCNFs and NIP-BCNFs from aqueous hemoglobin solutions were investigated in a batch system. The results showed that MIP-BCNFs are promising materials for purification of hemoglobin with high adsorption capacity, significant selectivity and reusability., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Identification of pathogenic microorganisms by traditional methods is slow and cumbersome. Therefore, the focus today is on developing new and quicker analytical methods. In this study, a Surface Plasmon Resonance (SPR) sensor with a microcontact imprinted sensor chip was developed for detecting Salmonella paratyphi . For this purpose, the stamps of the target microorganism were prepared and then, microcontact S. paratyphi -imprinted SPR chips were prepared with the functional monomer N-methacryloyl-L-histidine methyl ester (MAH). Characterization studies of the SPR chips were carried out with ellipsometry and scanning electron microscopy (SEM). The real-time Salmonella paratyphi detection was performed within the range of 2.5 × 10⁶-15 × 10⁶ CFU/mL. Selectivity of the prepared sensors was examined by using competing bacterial strains such as Escherichia coli , Staphylococcus aureus and Bacillus subtilis . The imprinting efficiency of the prepared sensor system was determined by evaluating the responses of the SPR chips prepared with both molecularly imprinted polymers (MIPs) and non-imprinted polymers (NIPs). Real sample experiments were performed with apple juice. The recognition of Salmonella paratyphi was achieved using these SPR sensor with a detection limit of 1.4 × 10⁶ CFU/mL. In conclusion, SPR sensor has the potential to serve as an excellent candidate for monitoring Salmonella paratyphi in food supplies or contaminated water and clearly makes it possible to develop rapid and appropriate control strategies., Competing Interests: The authors declare no conflict of interest.

Detection of protein C (PC) in human serum was performed by quartz crystal microbalance (QCM) based on molecular imprinting technique (MIP). The high-resolution and mass-sensitive QCM based sensor was integrated with high sensitivity and selectivity of the MIP technique. The PC microcontact imprinted (PC-μCIP) nanofilm was prepared on the glass surface. Then, the PC-μCIP/QCM sensor was prepared with 2-hydroxyethyl methacrylate (HEMA), ethylene glycol dimethacrylate (EGDMA) and N-methacryloyl l-histidine methylester (MAH) as the functional monomer with copper(II) ions. The polymerization was performed under UV light (100W and 365nm) for 20-25min under nitrogen atmosphere. The characterization studies of QCM sensor were done by observation using atomic force microscopy (AFM), contact angle measurements, ellipsometry and fourier transform infrared spectroscopy (FTIR). Detection of PC was investigated in a concentration range of 0.1-30μg/mL. Selectivity of PC-μCIP and PC non-imprinted/QCM (PC-non-μCIP) sensors for PC determination was investigated by using proteins namely hemoglobin (Hb), human serum albumin (HSA) and fibrinogen solutions. QCM sensor was also used for detection of PC molecules in aqueous solutions and human plasma. The detection limit was determined as 0.01μg/mL for PC analysis. The PC-μCIP/QCM sensor was used for five consecutive adsorption-desorption cycles. According to the results, the PC-μCIP/QCM sensor had obtained high selectivity and sensitivity for detection of PC molecules., (Copyright © 2016 Elsevier B.V. All rights reserved.)

In this study, thiophilic cryogels were prepared by two different approaches and they were used in purification of IgG from aqueous solutions and human plasma. In the first approach, poly(2-hydoxyethyl methacrylate) [PHEMA] cryogel disks were prepared. The PHEMA cryogel disks were activated by divinylsulfone (DVS) and 2-mercapto ethanol was attached. In the second approach, poly(2-hydroxyethyl methacrylate-etylene glycol dimethacrylate) [P(HEMA-EGDMA)] beads were synthesized and 2-mercapto ethanol was attached to the beads as a thiophilic ligand. In order to increase the surface area, P(HEMA-EGDMA)/PHEMA composite cryogel disks were prepared by embedding the P(HEMA-EGDMA) beads into PHEMA cryogels. Both the thiophilic PHEMA (T-PHEMA) cryogel disks and the thiophilic P(HEMA-EGDMA)/PHEMA [T-P(HEMA-EGDMA)/PHEMA] composite cryogel disks were characterized by Fourier transform infrared spectroscopy, surface area measurements, elemental analysis, swelling tests and scanning electron microscopy. The effects of salt concentration, pH, temperature, initial IgG concentration were analysed for IgG adsorption from aqueous solutions in batch mode. When lyotropic salt, Na2SO4, was used, the adsorption capacity was 27.5 mg/g and 68.7 mg/g for T-PHEMA and the T-P(HEMA-EGDMA)/PHEMA composite cryogel disks, respectively. 1.0 M NaCl was used as desorption agent. The change in adsorption capacity was not remarkable for a repetition of adsorption-desorption cycle ten times. Both the thiophilic cryogel disks and the thiophilic composite cryogel disks were also successfully used for IgG isolation from human plasma. The purity assayed by SDS-PAGE was 89%. The adsorption capacities were 74.8 mg/g and 137.4 mg/g in human plasma samples for the T-PHEMA cryogel disks and the T-P(HEMA-EGDMA)/PHEMA composite cryogel disks, respectively., (Copyright © 2013 Elsevier B.V. All rights reserved.)