scholarly journals Methodology and Applications of Disease Biomarker Identification in Human Serum

2007 ◽  
Vol 2 ◽  
pp. 117727190700200 ◽  
Author(s):  
Ziad J. Sahab ◽  
Suzan M. Semaan ◽  
Qing-Xiang Amy Sang
Keyword(s):  
Human Serum ◽  
Protein Separation ◽  
High Sensitivity ◽  
Disease Diagnosis ◽  
Plasma Lipoproteins ◽  
Great Promise ◽  
Protein Biomarkers ◽  
Serum Samples ◽  
Disease Biomarkers ◽  
Diagnosis And Prognosis

Biomarkers are biomolecules that serve as indicators of biological and pathological processes, or physiological and pharmacological responses to a drug treatment. Because of the high abundance of albumin and heterogeneity of plasma lipoproteins and glycoproteins, biomarkers are difficult to identify in human serum. Due to the clinical significance the identification of disease biomarkers in serum holds great promise for personalized medicine, especially for disease diagnosis and prognosis. This review summarizes some common and emerging proteomics techniques utilized in the separation of serum samples and identification of disease signatures. The practical application of each protein separation or identification technique is analyzed using specific examples. Biomarkers of cancers of prostate, breast, ovary, and lung in human serum have been reviewed, as well as those of heart disease, arthritis, asthma, and cystic fibrosis. Despite the advancement of technology few biomarkers have been approved by the Food and Drug Administration for disease diagnosis and prognosis due to the complexity of structure and function of protein biomarkers and lack of high sensitivity, specificity, and reproducibility for those putative biomarkers. The combination of different types of technologies and statistical analysis may provide more effective methods to identify and validate new disease biomarkers in blood.

scholarly journals Novel circulating protein biomarkers for thyroid cancer determined through data-independent acquisition mass spectrometry

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e9507
Author(s):  
Dandan Li ◽  
Jie Wu ◽  
Zhongjuan Liu ◽  
Ling Qiu ◽  
Yimin Zhang
Keyword(s):  
Mass Spectrometry ◽  
Area Under The Curve ◽  
High Sensitivity ◽  
Factor H ◽  
Complement Factor ◽  
Protein Biomarkers ◽  
Serum Samples ◽  
Data Independent Acquisition ◽  
Median Serum ◽  
And Control

Background Distinguishing between different types of thyroid cancers (TC) remains challenging in clinical laboratories. As different tumor types require different clinical interventions, it is necessary to establish new methods for accurate diagnosis of TC. Methods Proteomic analysis of the human serum was performed through data-independent acquisition mass spectrometry for 29 patients with TC (stages I–IV): 13 cases of papillary TC (PTC), 10 cases of medullary TC (MTC), and six cases follicular TC (FTC). In addition, 15 patients with benign thyroid nodules (TNs) and 10 healthy controls (HCs) were included in this study. Subsequently, 17 differentially expressed proteins were identified in 291 patients with TC, including 247 with PTC, 38 with MTC, and six with FTC, and 69 patients with benign TNs and 176 with HC, using enzyme-linked immunosorbent assays. Results In total, 517 proteins were detected in the serum samples using an Orbitrap Q-Exactive-plus mass spectrometer. The amyloid beta A4 protein, apolipoprotein A-IV, gelsolin, contactin-1, gamma-glutamyl hydrolase, and complement factor H-related protein 1 (CFHR1) were selected for further analysis. The median serum CFHR1 levels were significantly higher in the MTC and FTC groups than in the PTC and control groups (P < 0.001). CFHR1 exhibited higher diagnostic performance in distinguishing patients with MTC from those with PTC (P < 0.001), with a sensitivity of 100.0%, specificity of 85.08%, area under the curve of 0.93, and detection cut-off of 0.92 ng/mL. Conclusion CFHR1 may serve as a novel biomarker to distinguish PTC from MTC with high sensitivity and specificity.

Synthesis of novel cationic carbon dots and application to quantitative detection of K+ in human serum samples

2019 ◽  
Vol 43 (46) ◽  
pp. 17937-17940 ◽  
Author(s):  
Zideng Gao ◽  
Shunyi Wang ◽  
Zijun Xu ◽  
Jin Liu ◽  
Yuanfang Huang ◽  
...  
Keyword(s):  
Human Serum ◽  
Carbon Dots ◽  
High Sensitivity ◽  
Quantitative Detection ◽  
Serum Samples ◽  
Human Serum Samples

Novel cationic carbon dots were synthesized in a simple way and applied to detect K+ in human serum samples with ultra-high sensitivity.

scholarly journals Modified Potentiometric Screen-Printed Electrodes Based on Imprinting Character for Sodium Deoxycholate Determination

Biomolecules ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 251 ◽  
Author(s):  
Ayman H. Kamel ◽  
Samar Ezzat ◽  
Mona A. Ahmed ◽  
Abd El-Galil E. Amr ◽  
Abdulrahman A. Almehizia ◽  
...  
Keyword(s):  
Serum Albumin ◽  
Human Serum ◽  
Great Influence ◽  
High Sensitivity ◽  
Sodium Deoxycholate ◽  
Thermal Polymerization ◽  
Potentiometric Sensors ◽  
Serum Samples ◽  
And Gender ◽  
Screen Printed

Potentiometric sensors have a great influence on the determination of most various compounds in their matrices. Therefore, efficient and new sensors were introduced to measure sodium Deoxycholate (NaDC) as a bile acid salt. These sensors are based on NaDC imprinted polymer (MIP) as sensory element. The MIP beads were synthesized using thermal polymerization pathway, in which acrylamide (AAm), ethylene glycol dimethacrylate (EGDMA), NaDC, and benzoyl peroxide (BPO) were used as the functional monomer, cross-linker, template, and initiator, respectively. The proposed sensors were fabricated using a coated screen-printed platform and the sensing membrane was modified by single-walled carbon nanotubes (SWCNTs) as an ion-to-electron transducer. The sensors exhibited high sensitivity that reached 4.7 × 10−5 M of near-Nernestian slope (−60.1 ± 0.9 mV/decade, r2 = 0.999 (n= 5)). In addition, the sensors revealed high selectivity, long lifetime, high potential stability, and conductivity that ensure reproducible and accurate results over a long time. MIP characterization was performed using Fourier Transform-Infrared (FT-IR) and a scanning electron microscope (SEM). Regarding the interaction of NaDC with serum albumin (SA), albumin is determined in human serum samples as human serum albumin (HSA), which was collected from different volunteers of different ages and gender.

scholarly journals Reusable, Noninvasive, and Sensitive Fluorescence Enhanced ZnO-Nanorod-Based Microarrays for Quantitative Detection of AFP in Human Serum

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Saima Rafique ◽  
Farukh Kiyani ◽  
Sumbal Jawaid ◽  
Rubina Nasir ◽  
Mahmoosh Ahmad ◽  
...  
Keyword(s):  
Human Serum ◽  
Dynamic Range ◽  
Limit Of Detection ◽  
Zno Nanorods ◽  
Protein Microarrays ◽  
Great Promise ◽  
Quantitative Detection ◽  
Zno Nanorod ◽  
Serum Samples ◽  
Protein Assay

The fabrication of sensitive protein microarrays such as PCR used in DNA microarray is challenging due to lack of signal amplification. The development of microarrays is utilized to improve the sensitivity and limitations of detection towards primal cancer detection. The sensitivity is enhanced by the use of ZnO-nanorods and is investigated as a substrate which enhance the florescent signal to diagnose the hepatocellular carcinoma (HCC) at early stages. The substrate for deposition of ZnO-nanorods is prepared by the conventional chemical bath deposition method. The resultant highly dense ZnO-nanorods enhance the fluorescent signal 7.2 times as compared to the substrate without ZnO-nanorods. The microarray showed sensitivity of 1504.7 ng ml-1 and limit of detection of 0.1 pg ml-1 in wide dynamic range of 0.05 pg-10 μg ml-1 for alpha fetoprotein (AFP) detection in 10% human serum. This immunoassay was successfully applied for human serum samples to detect tumor marker with good recoveries. The ZnO-nanorod substrate is a simple protein microarray which showed a great promise for developing a low-cost, sensitive, and high-throughput protein assay platform for several applications in both fundamental research and clinical diagnosis.

scholarly journals A Fluorescence Inner-Filter Effect Based Sensing Platform for Turn-On Detection of Glutathione in Human Serum

Sensors ◽  
2019 ◽  
Vol 19 (2) ◽  
pp. 228 ◽  
Author(s):  
Shurong Tang ◽  
Xiuhua You ◽  
Quanhui Fang ◽  
Xin Li ◽  
Guangwen Li ◽  
...  
Keyword(s):  
Human Serum ◽  
Limit Of Detection ◽  
Cost Effective ◽  
Inner Filter Effect ◽  
Disease Diagnosis ◽  
Fluorescent Properties ◽  
Serum Samples ◽  
Turn On ◽  
Filter Effect ◽  
Sensing Platform

A novel turn-on fluorescence assay was developed for the rapid detection of glutathione (GSH) based on the inner-filter effect (IFE) and redox reaction. Molybdenum disulfide quantum dots (MoS2 QDs), which have stable fluorescent properties, were synthesized with hydrothermal method. Manganese dioxide nanosheets (MnO2 NSs) were prepared by exfoliating the bulk δ-MnO2 material in bovine serum albumin (BSA) aqueous solution. The morphology structures of the prepared nanoparticles were characterized by transmission electron microscope (TEM). Studies have shown that the fluorescence of MoS2 QDs could be quenched in the presence of MnO2 NSs as a result of the IFE, and is recovered after the addition of GSH to dissolve the MnO2 NSs. The fluorescence intensity showed a good linear relationship with the GSH concentration in the range 20–2500 μM, the limit of detection was 1.0 μM. The detection method was applied to the analysis of GSH in human serum samples. This simple, rapid, and cost-effective method has great potential in analyzing GSH and in disease diagnosis.

Simultaneous measurement of 13 circulating vitamin D3 and D2 mono and dihydroxy metabolites using liquid chromatography mass spectrometry

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Carl Jenkinson ◽  
Reena Desai ◽  
Andrzej T. Slominski ◽  
Robert C. Tuckey ◽  
Martin Hewison ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Vitamin D ◽  
Liquid Chromatography ◽  
Human Serum ◽  
High Sensitivity ◽  
Reference Level ◽  
Vitamin D Metabolites ◽  
Serum Samples ◽  
Human Serum Samples ◽  
Limit Of Quantitation

Abstract Objectives Clinical evaluation of vitamin D status is conventionally performed by measuring serum levels of a single vitamin D metabolite, 25-hydroxyvitamin D predominantly by immunoassay methodology. However, this neglects the complex metabolic pathways involved in vitamin D bioactivity, including two canonical forms D3 and D2, bioactive 1,25-dihydroxy metabolites and inactive 24-hydroxy and other metabolites. Methods Liquid chromatography-tandem mass spectrometry (LC-MS/MS) can measure multiple analytes in a sample during a single run with high sensitivity and reference level specificity. We therefore aimed to develop and validate a LC-MS/MS method to measure simultaneously 13 circulating vitamin D metabolites and apply it to 103 human serum samples. Results The LC-MS/MS method using a Cookson-type derivatization reagent phenyl-1,2,4-triazoline-3,5-dione (PTAD) quantifies 13 vitamin D metabolites, including mono and dihydroxy-metabolites, as well as CYP11A1-derived D3 and D2 metabolites in a single run. The lower limit of quantitation was 12.5 pg/mL for 1,25(OH)2D3 with accuracy verified by analysis of National Institute of Standards and Technology (NIST) 972a standards. Quantification of seven metabolites (25(OH)D3, 25(OH)D2, 3-epi-25(OH)D3, 20(OH)D3, 24,25(OH)2D3, 1,25(OH)2D3 and 1,20S(OH)2D3) was consistently achieved in human serum samples. Conclusions This profiling method can provide new insight into circulating vitamin D metabolite pathways forming the basis for improved understanding of the role of vitamin D in health and disease.

scholarly journals Multisite Comparison of High-Sensitivity Multiplex Cytokine Assays

2011 ◽  
Vol 18 (8) ◽  
pp. 1229-1242 ◽  
Author(s):  
Elizabeth Crabb Breen ◽  
Sandra M. Reynolds ◽  
Christopher Cox ◽  
Lisa P. Jacobson ◽  
Larry Magpantay ◽  
...  
Keyword(s):  
Plasma Concentrations ◽  
High Sensitivity ◽  
Small Sample ◽  
Great Promise ◽  
Serum Samples ◽  
Meso Scale Discovery ◽  
Necrosis Factor Alpha ◽  
Multiplex Assays ◽  
Long Term Study

ABSTRACTThe concentrations of cytokines in human serum and plasma can provide valuable information aboutin vivoimmune status, but low concentrations often require high-sensitivity assays to permit detection. The recent development of multiplex assays, which can measure multiple cytokines in one small sample, holds great promise, especially for studies in which limited volumes of stored serum or plasma are available. Four high-sensitivity cytokine multiplex assays on a Luminex (Bio-Rad, BioSource, Linco) or electrochemiluminescence (Meso Scale Discovery) platform were evaluated for their ability to detect circulating concentrations of 13 cytokines, as well as for laboratory and lot variability. Assays were performed in six different laboratories utilizing archived serum from HIV-uninfected and -infected subjects from the Multicenter AIDS Cohort Study (MACS) and the Women's Interagency HIV Study (WIHS) and commercial plasma samples spanning initial HIV viremia. In a majority of serum samples, interleukin-6 (IL-6), IL-8, IL-10, and tumor necrosis factor alpha were detectable with at least three kits, while IL-1β was clearly detected with only one kit. No single multiplex panel detected all cytokines, and there were highly significant differences (P< 0.001) between laboratories and/or lots with all kits. Nevertheless, the kits generally detected similar patterns of cytokine perturbation during primary HIV viremia. This multisite comparison suggests that current multiplex assays vary in their ability to measure serum and/or plasma concentrations of cytokines and may not be sufficiently reproducible for repeated determinations over a long-term study or in multiple laboratories but may be useful for longitudinal studies in which relative, rather than absolute, changes in cytokines are important.

scholarly journals Epitope Fishing in Digested Biological Samples

2018 ◽  
Author(s):  
Maren Levernæs ◽  
Bassem Farhat ◽  
Inger Oulie ◽  
Sazan S. Abdullah ◽  
Elisabeth Paus ◽  
...  
Keyword(s):  
Protein Extraction ◽  
High Sensitivity ◽  
University Hospital ◽  
Intact Protein ◽  
Protein Biomarkers ◽  
Serum Samples ◽  
Protein Levels ◽  
Norwegian Radium Hospital ◽  
Endogenous Protein ◽  
Sensitivity And Selectivity

<p>Immunocapture LC-MS/MS is a promising technique to ensure high sensitivity and selectivity of low-abundant protein biomarkers. For this purpose, the use of monoclonal antibodies (mAb) is especially attractive as they are renewable reagents that can be standardized. In this article we investigated the possibility of using mAbs developed against intact proteins to capture proteotypic epitope peptides. Three mAbs were tested, and all selectively extracted proteotypic epitope peptides from a complex sample. Compared to intact protein extraction, this concept which we call epitope fishing provided cleaner extracts, which further improved the sensitivity. Analysis of three patient samples demonstrated that epitope fishing can be used for the determination of different endogenous protein levels. </p><p><br></p><p> p.p1 {margin: 0.0px 0.0px 0.0px 0.0px; font: 14.5px Arial} </p><p>Human serum from healthy subjects was obtained from Oslo University Hospital, Ullevål (Oslo, Norway), and serum samples from cancer patients were supplied by the Norwegian Radium Hospital, Oslo University Hospital. All serum samples were stored at −30 °C. The use of patient samples for our research purposes was approved by the Norwegian Regional Committee for Medical Research Ethics (REK, http://helseforskning.etikkom.no). Informed consent was obtained from all subjects. Methods used to analyze all serum samples were in accordance with relevant guidelines and regulations.</p><br><p></p>

scholarly journals Serum MicroRNA Expression Profile as a Biomarker in the Diagnosis and Prognosis of Pancreatic Cancer

2012 ◽  
Vol 58 (3) ◽  
pp. 610-618 ◽  
Author(s):  
Rui Liu ◽  
Xi Chen ◽  
Yiqi Du ◽  
Weiyan Yao ◽  
Lin Shen ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
Expression Profile ◽  
High Sensitivity ◽  
Diagnostic Value ◽  
Serum Samples ◽  
Double Blind ◽  
Serum Mirna ◽  
Diagnosis And Prognosis ◽  
Serum Microrna ◽  
Specificity And Sensitivity

Abstract BACKGROUND Detection of pancreatic cancer (PaC), particularly at early stages, remains a great challenge owing to lack of specific biomarkers. We sought to identify a PaC-specific serum microRNA (miRNA) expression profile and test its specificity and sensitivity as a biomarker in the diagnosis and prognosis of PaC. METHODS We obtained serum samples from 197 PaC cases and 158 age- and sex-matched cancer-free controls. We screened the differentially expressed serum miRNAs with Illumina sequencing by synthesis technology using pooled serum samples followed by RT-qPCR validation of a large number of samples arranged in multiple stages. We used risk score analysis to evaluate the diagnostic value of the serum miRNA profiling system. To assess the serum miRNA–based biomarker accuracy in predicting PaC, we performed additional double-blind testing in 77 PaC cases and 52 controls and diagnostic classification in 55 cases with clinically suspected PaC. RESULTS After the selection and validation process, 7 miRNAs displayed significantly different expression levels in PaC compared with controls. This 7 miRNA–based biomarker had high sensitivity and specificity for distinguishing various stages of PaC from cancer-free controls and also accurately discriminated PaC patients from chronic pancreatitis (CP) patients. Among the 7 miRNAs, miR-21 levels in serum were significantly associated with overall PaC survival. The diagnostic accuracy rate of the 7-miRNA profile was 83.6% in correctly classifying 55 cases with clinically suspected PaC. CONCLUSIONS These data demonstrate that the 7 miRNA–based biomarker can serve as a novel noninvasive approach for PaC diagnosis and prognosis.

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