Analytical techniques for multiplex analysis of protein biomarkers

Introduction: The importance of biomarkers for pharmaceutical drug development and clinical diagnostics is more significant than ever in the current shift toward personalized medicine. Biomarkers have taken a central position either as companion markers to support drug development and patient selection, or as indicators aiming to detect the earliest perturbations indicative of disease, minimizing therapeutic intervention or even enabling disease reversal. Protein biomarkers are of particular interest given their central role in biochemical pathways. Hence, capabilities to analyze multiple protein biomarkers in one assay are highly interesting for biomedical research. Areas covered: We here review multiple methods that are suitable for robust, high throughput, standardized, and affordable analysis of protein biomarkers in a multiplex format. We describe innovative developments in immunoassays, the vanguard of methods in clinical laboratories, and mass spectrometry, increasingly implemented for protein biomarker analysis. Moreover, emerging techniques are discussed with potentially improved protein capture, separation, and detection that will further boost multiplex analyses. Expert commentary: The development of clinically applied multiplex protein biomarker assays is essential as multi-protein signatures provide more comprehensive information about biological systems than single biomarkers, leading to improved insights in mechanisms of disease, diagnostics, and the effect of personalized medicine

Analytical techniques for multiplex analysis of protein biomarkers

Introduction: The importance of biomarkers for pharmaceutical drug development and clinical diagnostics is more significant than ever in the current shift toward personalized medicine. Biomarkers have taken a central position either as companion markers to support drug development and patient selection, or as indicators aiming to detect the earliest perturbations indicative of disease, minimizing therapeutic intervention or even enabling disease reversal. Protein biomarkers are of particular interest given their central role in biochemical pathways. Hence, capabilities to analyze multiple protein biomarkers in one assay are highly interesting for biomedical research. Areas covered: We here review multiple methods that are suitable for robust, high throughput, standardized, and affordable analysis of protein biomarkers in a multiplex format. We describe innovative developments in immunoassays, the vanguard of methods in clinical laboratories, and mass spectrometry, increasingly implemented for protein biomarker analysis. Moreover, emerging techniques are discussed with potentially improved protein capture, separation, and detection that will further boost multiplex analyses. Expert commentary: The development of clinically applied multiplex protein biomarker assays is essential as multi-protein signatures provide more comprehensive information about biological systems than single biomarkers, leading to improved insights in mechanisms of disease, diagnostics, and the effect of personalized medicine.</div

An Electrochemical Biosensor Platform for Testing of Dehydroepiandrosterone 3-Sulfate (DHEA?S) as a Model for Doping Materials

2-s2.0-85071341127Endogenous steroids such as dehydroepiandrosterone (DHEA) and dehydroepiandrosterone 3-sulfate (DHEA?S) have commonly used as doping materials by athletes and to date novel techniques are needed for detection of these molecules. In this study, antibody-based electrochemical biosensor has developed for testing level of the DHEA?S. For this aim, gold surfaces were initially modified with cysteamine (Cys) and then, DHEA?S antibody was immobilized on the surface via glutaraldehyde (GA) as a crosslinking agent. The stepwise modification of electrode surface was monitored by using various electrochemical techniques such as cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Linear range was determined as 2.5–100 ng/mL DHEA?S using differential pulse voltammetry (DPV) technique, as well. Moreover, repeatability (±S.D.), coefficient of variation (%) and limit of detection (LOD) values were calculated as 0.033, 1.030 and 3.971, respectively. Also, DHEA?S in synthetic serum and urine samples were successfully determined with standard addition method and confirmation analysis were performed with liquid chromatography quadrupole-time of flight mass spectrometry (LC-QTOF/MS) system. The selectivity was studied with the addition of some interfering molecules (testosterone, bovine serum albumin (BSA), cholesterol, uric acid, lactic acid, codein (COD), ascorbic acid, DHEA). Consequently, this work is proposed as practical, innovative and cost-effective technique that can be easily adapted for the miniaturized form for the analysis of other doping substances as well as DHEA?S for the future works. © 2019 Wiley-VCH Verlag GmbH & Co. KGaA, WeinheimEge Üniversitesi: 18?EGEMATAL?001This study was supported by Ege University Scientific Research Project Coordination (BAP) coded 18?EGEMATAL?001. All experiments were conducted in Ege University Central Research Test and Analysis Laboratory Application and Research Center (EGE?MATAL). Also, throughout all experiments, facilities provided by 16?DPT?001 project were used

Catechol-Attached Polypeptide with Functional Groups as Electrochemical Sensing Platform for Synthetic Cannabinoids

Herein, we first constructed a functional surface using a catechol-attached polypeptide (CtP) for the detection of JWH-018 (N-4-hydroxypentyl metabolite). K2 antibody was then incorporated to the polymer via covalent cross-linker. Step-by-step modifications on the glassy carbon electrode surface were characterized by electrochemical measurements such as differential pulse voltammetry, cyclic voltammetry, impedance spectroscopy, and X-ray photoelectron spectroscopy. Linearity and the limit of detection for JWH-018 (N-4-hydroxypentyl metabolite) were determined as 10-500 ng/mL with an equation of y = 0.0018x + 0.136 (R2 = 0.993) and 5.892 ng/mL, respectively. The selectivity of the biosensor was evaluated with different interfering molecules (methamphetamine, codeine, and cocaine). Finally, the biosensor was successfully used in the determination of JWH-018 (N-4-hydroxypentyl metabolite) in spiked synthetic urine samples, and a high-performance liquid chromatography (HPLC) system was used as a reference method to confirm the sample application. The results show that this biosensor platform can be applied to detect other JWH series of synthetic cannabinoids with high sensitivity and accuracy. Copyright © 2019 American Chemical Society.2010K120810, 2016K121190; 117Z152The authors would like to thank the Turkish Scientific and Research Council (Project No. 117Z152) and Istanbul Technical University Research Fund for financial support. The Republic of Turkey, Ministry of Development, is also acknowledged (Project Grant Nos. 2016K121190 and 2010K120810) for providing the infrastructure to Ege University Central Research, Testing and Analysis Laboratory, Research and Application Center (EGE-MATAL)