A Comparison of Various Chips Used for the Manufacture of Biosensors Applied in Non-Fluidic Array SPRi, Based on the Example of Determination of Cathepsin D

Non-fluidic array SPR imaging (SPRi) with appropriate biosensors is a new tool for the determination of various biomarkers in body fluids. Numerous biomarkers can be determined without signal enhancement or preliminarily preconcentration. The introduction of a new material solution of the chip may increase the scope of the application of this technique. Solutions with adhesive separating foil and an Ag/Au chip were compared with the previously used two-paint separating polymer and pure gold chip. These solutions were tested using the example of a biosensor for cathepsin D (Cath D), which consisted of pepstatin A (a Cath D inhibitor) immobilized via a cysteamine linker using the NHS/EDC protocol. Four material versions of the Cath D biosensor proved adequate in terms of range of linearity, LOQ, precision and recovery. All four versions of the biosensor were used for the determination of Cath D in the blood serum patients with glioblastoma and control samples, producing very similar results and showing an elevated biomarker concentration in the case of cancer. Therefore, the problem of determining the correct level of Cath D in the serum of healthy individuals has been resolved, correcting literature data which ranged over three orders of magnitude.

Trends in the development of sensor devices based on surface plasmon resonance (Review)

Trends in the development of modern sensory devices based on surface plasmon resonance (SPR) are considered. The basic principles of construction of SPR sensor are given. For excitation of surface plasmons on the surface of sensitive elements of biosensory, a prism of total internal reflection is used or a dielectric substrate are used. A thin (dozens nm) film of high-conductive metal (mainly gold or silver) is applied to the working surface of the prisms or dielectric substrate. In a typical observation experiment, SPR is measured dependence on the angle of increasing light intensity, reflected by the resonance sensitive surface of the prism (chip). The optical schemes and principles of work of various SPR sensors are considered: - SPR Sensors with angular modulation, which are the most commonly used method based on the corner registration, in which the SPR occurs. The surface of the metal film is irradiated by monochromatic light and scans on a certain range of angles. There is a kind of SPR sensors with angular modulation, in which there is no mechanical scan of the angle of fall. Such sensors are entirely necessary for excitation of PPRs a set of angles is obtained due to a divergent or convergent light beam. - PPR sensors with a wavelength modulation is based on fixing an angle of falling light at a certain value and modulation of the wavelength of the incident light. Excitation of surface plasmons leads to a characteristic failure in the spectrum of reflected radiation. - Phase sensitive SPR sensors in which a change in the phase of the light wave associated with the surface plasma is measured on one corner of the fall and the wavelength of the light wave and is used as the output signal. - SPR imaging sensors in which the Technology of SPR imaging (SPRi) combines the sensitivity of the SPR with spatial image capabilities. The SPRI circuit uses as a fixed angle (as a rule, a slightly left angle of the SPR) and a fixed wavelength to measure changes in the reflection ability (Δ% R) that occur when the curve of the SPR is shifted due to the change in the refractive index above the surface of the sensor element. - SPR imaging sensors polarization contrast. In order to improve the quality of high-performance SPR imaging sensors in terms of sensitivity and resolution, the method of polarization contrast is used Disadvantages and advantages of SPR sensors are constructed with different principles are considered. The design and prospect of the use of achromatic and suburchast wave plates in the PPR imaging sensors with polarization contrast are considered.

Combination of Digital Image Processing and Statistical Data Segmentation to Enhance SPR and SPRi Sensor-responses

The work reports the combination of basics digital image processing (DIP) techniques and statistical segmentation strategy (SDS) to improve surface plasmon resonance curve (SPRc) and SPR imaging (SPRi) sensors' performance. The SPR image is used for sensing and monitoring biological events in the so-called SPR imaging process. In the traditional SPR process based on the attenuated total reflection (ATR) method, the image is used to create the SPR curve, and the curve features tracking is employed on sensing applications. The SPR curve features are enhanced after the pixels of the SPR image have been processed with lowcomplexity filters in the spatial domain (brightness, contrast, threshold, and morphological). The bootstrap was used as a statistical processing approach, selecting lines and columns from the image that was less affected by imperfections and noises in the image detector, and consequently reducing the SPR sensor instrumentation disturbances. Experimental tests with reversible binding water-mixture were performed, and both image and statistical processing were reported. The combination of DIP and SDS approaches improves the extraction of the curve features, increasing the performance in terms of resonance position sensitivity until 81%.

An Immunosensor for the Determination of Cathepsin S in Blood Plasma by Array SPRi—A Comparison of Analytical Properties of Silver–Gold and Pure Gold Chips

The array SPR imaging (SPRi) technique is well suited to the determination of biomarkers in body fluids, called liquid biopsy. No signal enhancement or analyte preconcentration is required. With the aim of achieving signal enhancement and lowering the cost of a single determination, the replacement of gold-covered chips by silver–gold chips was investigated. The aim of this work was to investigate the analytical characteristics of a biosensor formed on a Ag/Au chip and to compare them with those of a biosensor formed on a gold chip. A biosensor for the determination of cathepsin S (Cath S) was chosen as an example. The biosensor consisted of the linker cysteamine and an immobilized rat monoclonal antibody specific for cathepsin S. Both biosensors exhibited a Langmuirian response to Cath S concentration, with linear response ranging from LOQ to 1.5 ng mL−1. The LOQ is 0.1 ng mL−1 for the biosensor formed on the Ag/Au chip, and 0.22 ng mL−1 for that formed on the gold chip. Recoveries and precision for medium and high Cath S concentrations were acceptable for both biosensors, i.e., precision better than 10% and recoveries within the range 102–105%. However, the results for the lowest Cath S concentration were better for the biosensor formed on the Ag/Au chip (9.4 and 106% for precision and recovery, respectively). Generally, no significant differences in analytical characteristics were observed between the Ag/Au and Au chips. The two biosensors were also compared in the determination of Cath S in real samples. Nine plasma samples from healthy donors and nine from patients with ovarian cancer were analyzed for Cath S concentration with the biosensors formed on Ag/Au and Au chips. The results obtained with the two biosensors were very similar and show no significant differences on the Bland–Altman plot. The Cath S concentration in the blood plasma of ovarian cancer patients was elevated by one order of magnitude as compared with the control (12.6 ± 3.6 vs. 1.6 ± 1.2 ng mL−1).

Nanoparticle Determination in Water by LED-Excited Surface Plasmon Resonance Imaging

The increasing popularity of nanoparticles in many applications has led to the fact that these persistent materials pollute our environment and threaten our health. An online sensor system for monitoring the presence of nanoparticles in fresh water would be highly desired. We propose a label-free sensor based on SPR imaging. The sensitivity was enhanced by a factor of about 100 by improving the detector by using a high-resolution camera. This revealed that the light source also needed to be improved by using LED excitation instead of a laser light source. As a receptor, different self-assembled monolayers have been screened. It can be seen that the nanoparticle receptor interaction is of a complex nature. The best system when taking sensitivity as well as reversibility into account is given by a dodecanethiol monolayer on the gold sensor surface. Lanthanide-doped nanoparticles, 29 nm in diameter and with a similar refractive index to the most common silica nanoparticles were detected in water down to 1.5 µg mL−1. The sensor can be fully regenerated within one hour without the need for any washing buffer. This sensing concept is expected to be easily adapted for the detection of nanoparticles of different size, shape, and composition, and upon miniaturization, suitable for long-term applications to monitor the quality of water.

An SPR Imaging immunosensor for leptin determination in blood plasma

Leptin is a hormone that has a fundamental role in the regulation of feeding and energy balance. A developed specific SPRi immunosensor for leptin may be a new tool for...