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 Exploring peptide capture by anti-protein antibodies for LC-MS-based biomarker determination

2018 ◽  
Author(s):  
Maren Levernæs ◽  
Bassem Farhat ◽  
Inger Oulie ◽  
Sazan S. Abdullah ◽  
Elisabeth Paus ◽  
...  
Keyword(s):  
Protein Extraction ◽  
High Sensitivity ◽  
Intact Protein ◽  
Protein Biomarkers ◽  
Promising Technique ◽  
Intact Proteins ◽  
Protein Levels ◽  
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 (anti-protein antibodies) 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 peptide capture by anti-protein antibodies provided cleaner extracts, which further improved the sensitivity. Analysis of three patient samples demonstrated that p can be used for the determination of different endogenous protein levels. </p><p></p>

scholarly journals Exploring peptide capture by anti-protein antibodies for LC-MS-based biomarker determination

2018 ◽  
Author(s):  
Maren Levernæs ◽  
Bassem Farhat ◽  
Inger Oulie ◽  
Sazan S. Abdullah ◽  
Elisabeth Paus ◽  
...  
Keyword(s):  
Protein Extraction ◽  
High Sensitivity ◽  
Intact Protein ◽  
Protein Biomarkers ◽  
Promising Technique ◽  
Intact Proteins ◽  
Protein Levels ◽  
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 (anti-protein antibodies) 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 peptide capture by anti-protein antibodies provided cleaner extracts, which further improved the sensitivity. Analysis of three patient samples demonstrated that p can be used for the determination of different endogenous protein levels. </p><p></p>

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.

Protein biomarkers for multiple sclerosis: semi-quantitative analysis of cerebrospinal fluid candidate protein biomarkers in different forms of multiple sclerosis

2012 ◽  
Vol 18 (8) ◽  
pp. 1081-1091 ◽  
Author(s):  
Timucin Avsar ◽  
Didem Korkmaz ◽  
Melih Tütüncü ◽  
N Onat Demirci ◽  
Sabahattin Saip ◽  
...  
Keyword(s):  
Multiple Sclerosis ◽  
Cerebrospinal Fluid ◽  
Disease Process ◽  
Underlying Disease ◽  
Protein Biomarkers ◽  
Serum Samples ◽  
Neurofilament Light Chain ◽  
Neurofilament Light ◽  
Protein Levels ◽  
Clinical Subtype

Background: The complex pathogenesis of multiple sclerosis, combined with an unpredictable prognosis, requires identification of disease-specific diagnostic and prognostic biomarkers. Objective: To determine whether inflammatory proteins, such as neurofilament light chain, myelin oligodendrocyte glycoprotein and myelin basic protein, and neurodegenerative proteins, such as tau and glial fibrillary acidic protein, can serve as biomarkers for predicting the clinical subtype and prognosis of MS. Methods: Cerebrospinal fluid and serum samples were collected from patients with a diagnosis of clinically isolated syndrome ( n = 46), relapsing–remitting MS ( n = 67) or primary-progressive MS ( n = 22) along with controls having other non-inflammatory neurological disease ( n = 22). Western blot analyses were performed for the listed proteins. Protein levels were compared among different clinical subtypes using one-way analysis of variance analysis. The k-nearest neighbour algorithm was further used to assess the predictive use of these proteins for clinical subtype classification. Results: The results showed that each of tau, GFAP, MOG and NFL protein concentrations differed significantly ( p < 0.001) in multiple sclerosis clinical subtypes compared with the controls. Levels of the proteins also differed between the multiple sclerosis clinical subtypes, which may be associated with the underlying disease process. Classification studies revealed that these proteins might be useful for identifying multiple sclerosis clinical subtypes. Conclusions: We showed that select biomarkers may have potential in identifying multiple sclerosis clinical subtypes. We also showed that the predictive value of the prognosis increased when using a combination of the proteins versus using them individually.

scholarly journals Novel low-delta value troponin algorithms have good precision for rule-out of NSTE-ACS

2020 ◽  
Vol 41 (Supplement_2) ◽  
Author(s):  
H Tjora ◽  
O.T Steiro ◽  
J Langorgen ◽  
T Omland ◽  
P Collinson ◽  
...  
Keyword(s):  
False Negative ◽  
False Negative Rate ◽  
High Sensitivity ◽  
Public Institution ◽  
University Hospital ◽  
P Value ◽  
Funding Source ◽  
Good Precision ◽  
Serum Samples ◽  
Rule Out

Abstract Background Rapid rule-out algorithms for non-ST-elevation infarction (NSTEMI) may be beneficial for logistics in the emergency room. Current algorithms are designed to rule-out NSTEMI, but do not differentiate between unstable angina (UAP) in need of revascularization and non-cardiac chest pain patients (NCCP) who could be discharged. Recent improvements in analytical precision of high sensitivity troponin (cTn) assays allow for trialing algorithms with very small delta values. Purpose Could the use of lower delta values produce rule-out algorithms for NSTE-ACS with a false negative rate of ≤5%, and a sufficient high rule-out rate of patients with NCCP. Method 927 patients with suspected NSTE-ACS were consecutively included. Serum samples were collected at 0, 3 and 8–12 hours. The final diagnosis was adjudicated by two independent cardiologists based on all clinical data including routine cTnT. The 0- and 3-hour samples were additionally measured for cTnIand cTnI from Singulex Clarity System (cTnI(sgx)). The diagnostic performance to rule-out NSTE-ACS was compared between one low-delta value algorithm from each assay (cTnT, cTnI and cTnT). Results The prevalence of NSTEMI was 13.4%, UAP 11.4% and NCCP 60%. Median age was 63 years, 60% males. Fig. 1 shows baseline and 3-hour delta cTn values for the UAP and NCCP patients for the three different assays. The baseline cTn value differed significantly between UAP and NCCP for all assays, p value &lt;0.001. The novel low-delta cTnT algorithm (Table 1) ruled out 8 NSTE-ACS patients (3.5%), the cTnI algorithm and cTnI (sgx) algorithm ruled out 11 (4.8%) and 12 (5.2%) patients with NSTE-ACS, respectively. Moreover, the cTnT algorithm allocated 35.3% of the NCCP patients to discharge. Respective numbers for the cTnI(sgx) and cTnI algorithm were 30.6% and 33.5%. Comparing the ROC curves, the cTnT algorithm had significantly higher AUC compared to the cTnI(sgx) algorithm (p value =0.005, DeLong test). Conclusion The low-delta value algorithms correctly ruled in ≥95% of the NSTE-ACS patients whilst &gt;30% of NCCP patients were ruled out. The cTnT algorithm had the best performance with a significant higher AUC compared to the cTnI(sgx) algorithm. Figure 1 Funding Acknowledgement Type of funding source: Public Institution(s). Main funding source(s): Western Norway Regional Health Authority, Haukeland and Stavanger University hospital

Schottky diodes based on 2D materials for environmental gas monitoring: a review on emerging trends, recent developments and future perspectives

2021 ◽  
Author(s):  
Minu Mathew ◽  
Chandra Sekhar Rout
Keyword(s):  
Gas Sensing ◽  
2D Materials ◽  
Schottky Diodes ◽  
High Sensitivity ◽  
Future Perspectives ◽  
Working Temperature ◽  
Emerging Trends ◽  
Gas Sensing Properties ◽  
Recent Developments ◽  
Sensitivity And Selectivity

This review details the fundamentals, working principles and recent developments of Schottky junctions based on 2D materials to emphasize their improved gas sensing properties including low working temperature, high sensitivity, and selectivity.

Lanthanide coordination polymer-based biosensor for citrate detection in urine

2019 ◽  
Vol 11 (10) ◽  
pp. 1405-1409 ◽  
Author(s):  
Haoshuang Shen ◽  
Baoxia Liu ◽  
Daosheng Liu ◽  
Xu Zhu ◽  
Xiuhua Wei ◽  
...  
Keyword(s):  
Coordination Polymer ◽  
High Sensitivity ◽  
Lanthanide Coordination Polymer ◽  
Sensitivity And Selectivity

GMP ligand based LCP sensor for Cit with high sensitivity and selectivity was constructed.

Luminescent nucleotide/Tb3+ coordination polymer for Fe(ii) detection in human serum

2020 ◽  
Vol 12 (8) ◽  
pp. 1122-1130
Author(s):  
Yan Liu ◽  
Lu-Lu Peng ◽  
Wei-Xiong Huang ◽  
Hui-Zhuo Zhou ◽  
Li Xu
Keyword(s):  
Coordination Polymer ◽  
Human Serum ◽  
Adenosine Triphosphate ◽  
High Sensitivity ◽  
Sensitivity And Selectivity ◽  
Bridge Ligand

A luminescent nucleotide/Tb3+ coordination polymer (CP) was synthesized using adenosine triphosphate (ATP) molecules as a bridge ligand and Phen as a sensitizer, which showed high sensitivity and selectivity for Fe2+.

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