scholarly journals Advanced Techniques in Clinical Practice: Use of Lab-on-a-Chip Electrophoresis and Other Methods in Protein Profiling

2009 ◽  
Vol 28 (4) ◽  
pp. 274-278 ◽  
Author(s):  
Olgica Trenčevska ◽  
Vasko Aleksovski ◽  
Kiro Stojanoski
Keyword(s):  
Mass Spectrometry ◽  
Clinical Practice ◽  
Cystatin C ◽  
Biological Fluids ◽  
Neurological Diseases ◽  
Protein Profile ◽  
Lab On A Chip ◽  
Protein Profiling ◽  
Complete Analysis ◽  
Potential Biomarker

Advanced Techniques in Clinical Practice: Use of Lab-on-a-Chip Electrophoresis and Other Methods in Protein ProfilingProteins in clinical practice are analyzed as important parameters in the determination and treatment of different diseases. The scopes of the analyses are mainly concentrated in two levels - analyses of the complete protein profile, or determination of an isolated protein. In this work, despite of the use of conventional methods, mainly electrophoresis, new techniques have been implemented in protein analyses. Lab-on-a-chip is an electrophoretic technique that, when optimized, provides analyses of the total protein profile. When normal samples are compared to samples obtained from patients with different neurological diseases, characteristic patterns can be noted. Also, correlation and comparison can be made between the newly developed microchip electrophoresis method and the results obtained using the conventional techniques. When an analysis of a specific protein is necessary, mass spectrometry has proven to give best results, in both the se lectivity and specificity of analyses. It is believed that cystatin C is a potential biomarker in neurological diseases; therefore, the mass spectrometry method has been developed in order to obtain qualitative and quantitative analyses of biological fluids. Using the developed method of mass spectrometry immunoassay (MSIA), cystatin C was easily isolated and analyzed, obtaining complete analysis within minutes. The resulting mass spectra revealed various levels of cystatin C isoforms in serum and CSF samples.

Overview of the Chromatographic and Mass Spectrometry Analytical Methods for Determination of Lamivudine in Biological Fluids

2019 ◽  
Vol 15 (2) ◽  
pp. 103-108
Author(s):  
Xuwang Chen ◽  
Fanlong Bu ◽  
Rong Li ◽  
Guiyan Yuan ◽  
Yanyan Wang ◽  
...  
Keyword(s):  
United States ◽  
Mass Spectrometry ◽  
Clinical Practice ◽  
Analytical Methods ◽  
Review Paper ◽  
Biological Fluids ◽  
Animal Experiments ◽  
The United States ◽  
Therapeutic Drug

Background: Lamivudine was approved by Food and Drug Administration of the United States for the treatment of both HIV and HBV infection, which has been widely used as monotherapy or a component of combination therapy in clinics in many countries and nationalities. Methods: In this paper, the recent chromatographic and mass spectrometry analytical methods for the determination of lamivudine individually or combination with other drugs simultaneously were presented. These methods were widely applied in pharmacokinetics studies, bioequivalence studies, therapeutic drug monitoring studies, cell and animal experiments. Conclusion: The review paper might provide references for determining lamivudine in biological fluids, the intracorporal process of lamivudine, and the clinical practice by monitoring plasma concentration of lamivudine in the future.

Serum Protein Profiling with Mass Spectrometry for the Diagnosis of Myelodysplastic Syndromes.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 2362-2362
Author(s):  
Manuel Aivado ◽  
Dimitrios Spentzos ◽  
Ulrich Germing ◽  
Gil Alterovitz ◽  
Xiao-Ying Meng ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Bone Marrow ◽  
Serum Protein ◽  
Protein Profile ◽  
Protein Profiling ◽  
Protein Profiles ◽  
K Nearest Neighbor ◽  
Independent Validation ◽  
Validation Set ◽  
Tof Ms

Abstract Surface enhanced laser desorption/ionization-time of flight mass spectrometry (SELDI-TOF MS) has facilitated disease-specific serum protein profiles, which may become instrumental for diseases that are difficult to diagnose. The diagnosis of MDS can be very difficult since bone marrow dysplasia and peripheral cytopenia, the hallmarks of MDS, can be observed due to many reasons other than MDS. Moreover, cytomorphological evaluation of dysplasia requires extensive experience and is thus dependent on the examiner. Between 2–16% of the patients have a fibrotic bone marrow, which can impede appropriate bone marrow aspiration and smear. Finally, while cytogenetic examination can strongly support the diagnosis of MDS, only 40–50% of the patients have an abnormal karyotype. In the present study, we employed a SELDI-TOF MS-based procedure termed Pattern Track™ in order to analyze a total of 218 serum samples. We generated serum protein profiles from a first sample set comprising 74 patients with MDS, 39 control patients with cytopenia for reasons other than MDS, and 24 healthy persons. We fractionated their serum by means of anion exchange chromatography and applied the resulting serum fractions to weak cationic exchange as well as to reversed phase chromatography ProteinChip™ arrays. We randomly split this dataset into a learning (n=72) and a first independent validation set (n=41). Then, we used a k-nearest-neighbor algorithm to build a class predicting profile that consisted of 81 protein peaks. That profile was tested by leave-one-out cross validation and predicted the diagnosis MDS with an accuracy of 81.9% in the learning set (Fisher’s test, P=0.0000003). Then, we tested the profile on our first independent validation set and obtained a similar accuracy of 80.5% (P=0.0002). Its diagnostic performance and long-term reproducibility were confirmed by successfully applying it to a prospectively collected second independent validation set consisting of 81 new samples (P=0.0000006). Eventually, following serial chromatography, 1D gel electrophoresis, and tryptic peptide fingerprinting, we discovered the identity of 2 members of the profile. We conclude that our predicting serum protein profile represents a novel, non-invasive aid in distinguishing patients with MDS from patients with cytopenia for reasons other than MDS.

scholarly journals MALDI-ToF Protein Profiling as Potential Rapid Diagnostic Platform for COVID-19

2021 ◽  
Author(s):  
Prajkta Chivte ◽  
Zane LaCasse ◽  
Ventaka Devesh Reddy Seethi ◽  
Pratool Bharti ◽  
Elizabeth R. Gaillard
Keyword(s):  
Mass Spectrometry ◽  
High Specificity ◽  
Diagnostic Methods ◽  
Protein Profiling ◽  
Rt Pcr ◽  
High Demand ◽  
Non Invasive ◽  
Maldi Tof ◽  
Potential Biomarker ◽  
High Consistency

More than a year after the COVID-19 pandemic has been declared, the need still exists for accurate, rapid, inexpensive and non-invasive diagnostic methods that yield high specificity towards the current and newly emerging SARS-CoV-2 strains. Several studies have since established saliva as a more amenable specimen type in early detection and viral load quantitation as compared to the nasopharyngeal swabs. Considering the limitations and high demand for COVID-19 testing, we have employed MALDI-ToF mass spectrometry for the analysis of 60 gargle samples from human donors and compared the spectra with their respective RT-PCR results. Several standards including isolated human serum immunoglobulins and controls such as pre-COVID-19 saliva and heat inactivated SARS-CoV-2 virus were simultaneously analyzed to provide a relative view of the saliva and viral proteome as they would appear in this works methodology. Five potential biomarker peaks were established that demonstrated high consistency with PCR positive samples. Overall, the agreement of these results with RT-PCR testing was no less than 90% for the studied cohort, which consisted of young and largely asymptomatic student athletes. Further investigation of the potential biomarker peaks is necessary, however, from a clinical standpoint, these results are promising for a rapid and inexpensive COVID-19 assay.

The miR-21 potential of serving as a biomarker for liver diseases in clinical practice

2020 ◽  
Vol 48 (5) ◽  
pp. 2295-2305
Author(s):  
Jiawei Zhang ◽  
Dandan Li ◽  
Rui Zhang ◽  
Peng Gao ◽  
Rongxue Peng ◽  
...  
Keyword(s):  
Clinical Practice ◽  
Liver Diseases ◽  
Hepatic Disease ◽  
Noninvasive Detection ◽  
Diagnosis And Prognosis ◽  
Expected Performance ◽  
Potential Biomarker ◽  
Disease Condition ◽  
Complex Regulatory Network

The role of miR-21 in the pathogenesis of various liver diseases, together with the possibility of detecting microRNA in the circulation, makes miR-21 a potential biomarker for noninvasive detection. In this review, we summarize the potential utility of extracellular miR-21 in the clinical management of hepatic disease patients and compared it with the current clinical practice. MiR-21 shows screening and prognostic value for liver cancer. In liver cirrhosis, miR-21 may serve as a biomarker for the differentiating diagnosis and prognosis. MiR-21 is also a potential biomarker for the severity of hepatitis. We elucidate the disease condition under which miR-21 testing can reach the expected performance. Though miR-21 is a key regulator of liver diseases, microRNAs coordinate with each other in the complex regulatory network. As a result, the performance of miR-21 is better when combined with other microRNAs or classical biomarkers under certain clinical circumstances.

Suzuki–Miyaura cross-coupling for chemoproteomic applications

2020 ◽  
Author(s):  
Jian Cao ◽  
Ernest Armenta ◽  
Lisa Boatner ◽  
Heta Desai ◽  
Neil Chan ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Cross Coupling ◽  
Protein Profiling ◽  
Protein Labeling ◽  
Caspase 8 ◽  
Complex Cell ◽  
Bioorthogonal Chemistry ◽  
Reaction Conditions ◽  
Target Deconvolution ◽  
Palladium Catalyzed

Bioorthogonal chemistry is a mainstay of chemoproteomic sample preparation workflows. While numerous transformations are now available, chemoproteomic studies still rely overwhelmingly on copper-catalyzed azide –alkyne cycloaddition (CuAAC) or 'click' chemistry. Here we demonstrate that gel-based activity-based protein profiling (ABPP) and mass-spectrometry-based chemoproteomic profiling can be conducted using Suzuki–Miyaura cross-coupling. We identify reaction conditions that proceed in complex cell lysates and find that Suzuki –Miyaura cross-coupling and CuAAC yield comparable chemoproteomic coverage. Importantly, Suzuki–Miyaura is also compatible with chemoproteomic target deconvolution, as demonstrated using structurally matched probes tailored to react with the cysteine protease caspase-8. Uniquely enabled by the observed orthogonality of palladium-catalyzed cross-coupling and CuAAC, we combine both reactions to achieve dual protein labeling.

A Review of Analytical Methods for the Determination of Tibolone: Pharmacokinetics and Pharmaceutical Formulations Analysis and Application in Doping Control

2020 ◽  
Vol 17 (1) ◽  
pp. 31-39
Author(s):  
Marilene Lopes Ângelo ◽  
Fernanda de Lima Moreira ◽  
Ana Laura Araújo Santos ◽  
Hérida Regina Nunes Salgado ◽  
Magali Benjamim de Araújo
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Analytical Methods ◽  
Estrogenic Activity ◽  
Biological Fluids ◽  
Pharmaceutical Formulations ◽  
Doping Control ◽  
Specific Effects ◽  
In Doping

Background:: Tibolone is a synthetic steroid commercialized by Organon under the brand name Livial (Org OD14), which is used in hormone therapy for menopause management and treatment of postmenopausal osteoporosis. Tibolone is defined as a selective tissue estrogenic activity regulator (STEAR) demonstrating tissue-specific effects on several organs such as brain, breast, urogenital tract, endometrium, bone and cardiovascular system. Aims:: This work aims to (1) present an overview of important published literature on existing methods for the analysis of tibolone and/or its metabolites in pharmaceutical formulations and biological fluids and (2) to conduct a critical comparison of the analytical methods used in doping control, pharmacokinetics and pharmaceutical formulations analysis of tibolone and its metabolites. Results and conclusions: : The major analytical method described for the analysis of tibolone in pharmaceutical formulations is High Pressure Liquid Chromatography (HPLC) coupled with ultraviolet (UV) detection, while Liquid Chromatography (LC) or Gas Chromatography (GC) used in combination with Mass Spectrometry (MS) or tandem mass spectrometry (MS/MS) is employed for the analysis of tibolone and/or its metabolites in biological fluids.

scholarly journals Metabolic Profiling of Pitaya (Hylocereus polyrhizus) during Fruit Development and Maturation

Molecules ◽  
2019 ◽  
Vol 24 (6) ◽  
pp. 1114 ◽  
Author(s):  
Yawei Wu ◽  
Juan Xu ◽  
Yizhong He ◽  
Meiyan Shi ◽  
Xiumei Han ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Metabolic Profiling ◽  
Developmental Stages ◽  
Principal Component ◽  
Nutrient Content ◽  
Flight Mass Spectrometry ◽  
Potential Biomarker ◽  
Fruit Samples ◽  
Liquid Chromatography Tandem Mass ◽  
Hylocereus Polyrhizus

Pitaya (Hylocereus polyrhizus) has attracted much interest from consumers as it is a novelty fruit with high nutrient content and a tolerance to drought stress. As a group of attractive pigment- and health-promoting natural compounds, betalains represent a visual feature for pitaya fruit quality. However, little information on the correlation between betalains and relevant metabolites exists so far. Currently, color (Commission International del’Eclairage, CIE) parameters, betalain contents, and untargeted metabolic profiling (gas chromatography-time-of-flight-mass spectrometry, GC–MS and liquid chromatography tandem mass spectrometry, LC–MS) have been examined on ‘Zihonglong’ fruits at nine different developmental stages, and the variation character of the metabolite contents was simultaneously investigated between peel and pulp. Furthermore, principal component analysis (PCA) and partial least-squares discriminant analysis (PLS-DA) were used to explore metabolite profiles from the fruit samples. Our results demonstrated that the decrease of amino acid, accompanied by the increase of sugars and organic acid, might contribute to the formation of betalains. Notably, as one of four potential biomarker metabolites, citramalic acid might be related to betalain formation.

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