Proteins in biological fluids (blood, urine, cerebrospinal fluid) are important biomarkers of various pathological conditions. Protein biomarkers detection and quantification have been proven to be an indispensable diagnostic tool in clinical practice. There is a growing tendency towards using portable diagnostic biosensor devices for point-of-care (POC) analysis based on microfluidic technology as an alternative to conventional laboratory protein assays. In contrast to universally accepted analytical methods involving protein labeling, label-free approaches often allow the development of biosensors with minimal requirements for sample preparation by omitting expensive labelling reagents. The aim of the present work is to review the variety of physical label-free techniques of protein detection and characterization which are suitable for application in micro-fluidic structures and analyze the technological and material aspects of label-free biosensors that implement these methods. The most widely used optical and impedance spectroscopy techniques: absorption, fluorescence, surface plasmon resonance, Raman scattering, and interferometry, as well as new trends in photonics are reviewed. The challenges of materials selection, surfaces tailoring in microfluidic structures, and enhancement of the sensitivity and miniaturization of biosensor systems are discussed. The review provides an overview for current advances and future trends in microfluidics integrated technologies for label-free protein biomarkers detection and discusses existing challenges and a way towards novel solutions.

Diagnostic devices for point-of-care (POC) urine analysis (urinalysis) based on microfluidic technology have been actively developing for several decades as an alternative to laboratory based biochemical assays. Urine proteins (albumin, immunoglobulins, uromodulin, haemoglobin etc.) are important biomarkers of various pathological conditions and should be selectively detected by urinalysis sensors. The challenge is a determination of different oligomeric forms of the same protein, e.g., uromodulin, which have similar bio-chemical affinity but different physical properties. For the selective detection of different types of proteins, we propose to use a shear bulk acoustic resonator sensor with an additional electrode on the upper part of the bioliquid-filled channel for protein electric field manipulation. It causes modulation of the protein concentration over time in the near-surface region of the acoustic sensor, that allows to distinguish proteins based on their differences in diffusion coefficients (or sizes) and zeta-potentials. Moreover, in order to improve the sensitivity to density, we propose to use structured sensor interface. A numerical study of this approach for the detection of proteins was carried out using the example of albumin, immunoglobulin, and oligomeric forms of uromodulin in model urine solutions. In this contribution we prove the proposed concept with numerical studies for the detection of albumin, immunoglobulin, and oligomeric forms of uromodulin in urine models.

Point-of-care testing (POCT) devices are becoming increasingly popular in the medical community as an alternative to conventional laboratory testing, especially for home treatments or other forms of outpatient care. Multiple-use chemical sensors with minimal requirements for disposables are among the most practical and cost-effective POC diagnostic instruments, especially in managing chronic conditions. An affordable, simple, and easy-to-use optical sensor based on fast protein liquid chromatography with direct UV absorption detection was developed for the rapid determination of the total protein concentration in effluent peritoneal dialysate and for the assessment of protein losses in end-stage renal disease (ESRD) patients on constant ambulatory peritoneal dialysis (CAPD). The sensor employs non-disposable PD-10 desalting columns for the separation of molecules with different molecular weights and a deep UV LED (maximum at 285 nm) as a light source for optical detection. The analytic procedure is relatively simple, takes 10–15 min, and potentially can be performed by patients themselves or nursing staff without laboratory training. Preliminary clinical trials on a group of 23 patients on CAPD revealed a good concordance between the protein concentrations in dialysate samples measured with the sensor and an automated biochemical analyzer; the mean relative error was about 10%, which is comparable with routine clinical laboratory methods.

Milk is a product that requires quality control at all stages of production: from the dairy farm, processing at the dairy plant to finished products. Milk is a complex multiphase polydisperse system, whose components not only determine the quality and price of raw milk, but also reflect the physiological state of the herd. Today’s production volumes and rates require simple, fast, cost-effective, and accurate analytical methods, and most manufacturers want to move away from methods that use reagents that increase analysis time and move to rapid analysis methods. The review presents methods for the rapid determination of the main components of milk, examines their advantages and disadvantages. Optical spectroscopy is a fast, non-destructive, precise, and reliable tool for determination of the main constituents and common adulterants in milk. While mid-infrared spectroscopy is a well-established off-line laboratory technique for the routine quality control of milk, near-infrared technologies provide relatively low-cost and robust solutions suitable for on-site and in-line applications on milking farms and dairy production facilities. Other techniques, discussed in this review, including Raman spectroscopy, atomic spectroscopy, molecular fluorescence spectroscopy, are also used for milk analysis but much less extensively. Acoustic methods are also suitable for non-destructive on-line analysis of milk. Acoustic characterization can provide information on fat content, particle size distribution of fat and proteins, changes in the biophysical properties of milk over time, the content of specific proteins and pollutants. The basic principles of ultrasonic techniques, including transmission, pulse-echo, interferometer, and microbalance approaches, are briefly described and milk parameters measured with their help, including frequency ranges and measurement accuracy, are given.

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