Review: Detection and quantification of proteins in human urine

Sultan Aitekenov, Abduzhappar Gaipov, Rostislav Bukasov, Sultan Aitekenov, Abduzhappar Gaipov, Rostislav Bukasov

Abstract

Extensive medical research showed that patients, with high protein concentration in urine, have various kinds of kidney diseases, referred to as proteinuria. Urinary protein biomarkers are useful for diagnosis of many health conditions - kidney and cardio vascular diseases, cancers, diabetes, infections. This review focuses on the instrumental quantification (electrophoresis, chromatography, immunoassays, mass spectrometry, fluorescence spectroscopy, the infrared spectroscopy, and Raman spectroscopy) of proteins (the most of all albumin) in human urine matrix. Different techniques provide unique information on what constituents of the urine are. Due to complex nature of urine, a separation step by electrophoresis or chromatography are often used for proteomics study of urine. Mass spectrometry is a powerful tool for the discovery and the analysis of biomarkers in urine, however, costs of the analysis are high, especially for quantitative analysis. Immunoassays, which often come with fluorescence detection, are major qualitative and quantitative tools in clinical analysis. While Infrared and Raman spectroscopies do not give extensive information about urine, they could become important tools for the routine clinical diagnostics of kidney problems, due to rapidness and low-cost. Thus, it is important to review all the applicable techniques and methods related to urine analysis. In this review, a brief overview of each technique's principle is introduced. Where applicable, research papers about protein determination in urine are summarized with the main figures of merits, such as the limit of detection, the detectable range, recovery and accuracy, when available.

Keywords: Biomarkers; Fluorescence spectroscopy; Human serum albumin; Immunoassays; Mass spectrometry; Urine proteomics.

Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Copyright © 2020 The Author(s). Published by Elsevier B.V. All rights reserved.

Figures

Graphical abstract
Graphical abstract
Fig. 1
Fig. 1
Diagram of the GELFrEE device [68]. A gel column is utilized to achieve electrophoretic separation of proteins, analogous to SDS–PAGE, which are then eluted into the liquid-phase for manual collection. The fractionation can then be visualized by running a portion of the fractions on a SDS–PAGE gel. Reprinted from Refs. [95].
Fig. 2
Fig. 2
Schematic diagram for the competitive immunoassays. Reprinted from Ref. [109].
Fig. 3
Fig. 3
Schematic diagram for the non-competitive immunoassay. Reprinted from Ref. [109].
Fig. 4
Fig. 4
Schematic of graphene oxide-mediated fluorescence quenching aptasensor for the detection of albuminuria in urine and HSA in human serum. When albumin was added to the complex GO with the fluorescence-labeled aptamer, the aptamer detached from the complex to bind albumin, which resulted by an increase in fluorescence intensity. Reprinted from Refs. [132].
Fig. 5
Fig. 5
Raman spectra of PD patient urine and spent dialysate.(A) Averaged, baselined, and vector normalized Raman spectra from 362 urine specimens obtained from patients receiving PD therapy for ESKD. (B) Averaged, baselined, and vector normalized Raman spectra from 395 spent PD dialysate specimens. Reprinted from Ref. [164].

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