Journal of Mass Spectrometry and Advances in the Clinical Lab最新文献

{"title":"Development and validation of a multiplexed LC-MS/MS ketone body assay for clinical diagnostics","authors":"Robin H.J. Kemperman ,&nbsp;Rebecca D. Ganetzky ,&nbsp;Stephen R. Master","doi":"10.1016/j.jmsacl.2024.01.004","DOIUrl":"10.1016/j.jmsacl.2024.01.004","url":null,"abstract":"<div><h3>Objectives</h3><p>Ketone bodies (KBs) serve as important energy sources that spare glucose, providing the primary energy for cardiac muscle, skeletal muscle during aerobic exercise, and the brain during periods of catabolism. The levels and relationships between the KBs are critical indicators of metabolic health and disease. However, challenges in separating isomeric KBs and concerns about sample stability have previously limited their clinical measurement.</p></div><div><h3>Methods</h3><p>A novel 6.5-minute liquid chromatography-mass spectrometry-based assay was developed, enabling the precise measurement of alpha-, beta- and gamma-hydroxybutyrate, beta-hydroxyisobutyrate, and acetoacetate. This method was fully validated for human serum and plasma samples by investigating extraction efficiency, matrix effects, accuracy, recovery, intra- and inter-precision, linearity, lower limit of quantitation (LLOQ), carryover, specificity, stability, and more. From 107 normal samples, reference ranges were established for all analytes and the beta-hydroxybutyrate/acetoacetate ratio.</p></div><div><h3>Results</h3><p>All five analytes were adequately separated chromatographically. An extraction efficiency between 80 and 120 % was observed for all KBs. Accuracy was evaluated through spike and recovery using 10 random patient samples, with an average recovery of 85–115 % for all KBs and a coefficient of variation of ≤ 3 %. Coefficients of variation for intra- and inter-day imprecision were &lt; 5 %, and the total imprecision was &lt; 10 %. No significant interferences were observed. Specimens remained stable for up to 6 h on ice or 2 h at room temperature.</p></div><div><h3>Conclusions</h3><p>The developed method is highly sensitive and robust. It has been validated for use with human serum and plasma, overcoming stability concerns and providing a reliable and efficient quantitative estimation of ketone bodies.</p></div>","PeriodicalId":52406,"journal":{"name":"Journal of Mass Spectrometry and Advances in the Clinical Lab","volume":"31 ","pages":"Pages 49-58"},"PeriodicalIF":2.2,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2667145X2400004X/pdfft?md5=ecb251e99bcd8539ebb37c657c813fcf&pid=1-s2.0-S2667145X2400004X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139639230","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A high-throughput LC-MS/MS assay for piperaquine from dried blood spots: Improving malaria treatment in resource-limited settings","authors":"Daniel Blessborn ,&nbsp;Natpapat Kaewkhao ,&nbsp;Joel Tarning","doi":"10.1016/j.jmsacl.2023.12.004","DOIUrl":"https://doi.org/10.1016/j.jmsacl.2023.12.004","url":null,"abstract":"<div><h3>Background</h3><p>Malaria is a parasitic disease that affects many of the poorest economies, resulting in approximately 241 million clinical episodes and 627,000 deaths annually. Piperaquine, when administered with dihydroartemisinin, is an effective drug against the disease. Drug concentration measurements taken on day 7 after treatment initiation have been shown to be a good predictor of therapeutic success with piperaquine. A simple capillary blood collection technique, where blood is dried onto filter paper, is especially suitable for drug studies in remote areas or resource-limited settings or when taking samples from children, toddlers, and infants.</p></div><div><h3>Methods</h3><p>Three 3.2 mm discs were punched out from a dried blood spot (DBS) and then extracted in a 96-well plate using solid phase extraction on a fully automated liquid handling system. The analysis was performed using LC-MS/MS with a calibration range of 3 – 1000 ng/mL.</p></div><div><h3>Results</h3><p>The recovery rate was approximately 54–72 %, and the relative standard deviation was below 9 % for low, middle and high quality control levels. The LC-MS/MS quantification limit of 3 ng/mL is sensitive enough to detect piperaquine for up to 4–8 weeks after drug administration, which is crucial when evaluating recrudescence and drug resistance development. While different hematocrit levels can affect DBS drug measurements, the effect was minimal for piperaquine.</p></div><div><h3>Conclusion</h3><p>A sensitive LC-MS/MS method, in combination with fully automated extraction in a 96-well plate format, was developed and validated for the quantification of piperaquine in DBS. The assay was implemented in a bioanalytical laboratory for processing large-scale clinical trial samples.</p></div>","PeriodicalId":52406,"journal":{"name":"Journal of Mass Spectrometry and Advances in the Clinical Lab","volume":"31 ","pages":"Pages 19-26"},"PeriodicalIF":2.2,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2667145X23000421/pdfft?md5=cdb560cd35ba8072bfcd7a8f31c71e9d&pid=1-s2.0-S2667145X23000421-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139099707","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An LC-MS/MS-based platform for the quantification of multiple amyloid beta peptides in surrogate cerebrospinal fluid","authors":"Merve Oztug,&nbsp;Bilgin Vatansever,&nbsp;Gonca Altin,&nbsp;Muslum Akgoz,&nbsp;Suleyman Z. Can","doi":"10.1016/j.jmsacl.2024.01.002","DOIUrl":"https://doi.org/10.1016/j.jmsacl.2024.01.002","url":null,"abstract":"<div><h3>Introduction</h3><p>The accurate quantification of amyloid beta (Aβ) peptides in cerebrospinal fluid (CSF) is crucial for Alzheimer's disease (AD) research, particularly in terms of preclinical and biomarker studies. Traditional methods, such as the enzyme-linked immunosorbent assay (ELISA), have limitations. These include high costs, labor intensity, lengthy processes, and the possibility of cross-reactivity.</p></div><div><h3>Objectives</h3><p>The primary objectives of this research were twofold: to comprehensively characterize Aβ peptides and to develop a reliable and accurate method for the simultaneous quantification of Aβ 1–40 and Aβ 1–42 peptides in surrogate CSF that is traceable to the International System of Units (SI).</p></div><div><h3>Methods</h3><p>We developed a novel method that combined solid phase extraction (SPE) with isotope dilution liquid chromatography/tandem mass spectrometry (ID-LC/MSMS). SPE was employed to efficiently eliminate matrix interferences, while [15N] Aβ1-40 and [15N] Aβ1-42 served as internal standards to improve accuracy. In addition, we introduced Peptide Impurity Corrected Amino Acid Analysis (PICAA) to ensure traceability to the SI and reliable quantification of Aβ peptides.</p></div><div><h3>Results</h3><p>The developed platform demonstrated a linear calibration range of 300–20000 pg/ml for both Aβ1-42 and Aβ1-40 peptides, accompanied by strong correlation coefficients greater than 0.995. Quality Control (QC) samples demonstrated an accuracy of at least 90.0 %.</p></div><div><h3>Conclusion</h3><p>The enhanced specificity and flexibility of the developed platform potentially have implications for Alzheimer's disease diagnosis and future investigations of novel Aβ peptide biomarkers.</p></div>","PeriodicalId":52406,"journal":{"name":"Journal of Mass Spectrometry and Advances in the Clinical Lab","volume":"31 ","pages":"Pages 40-48"},"PeriodicalIF":2.2,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2667145X24000026/pdfft?md5=5be4e78996c7c16238efa4dcec36f6b3&pid=1-s2.0-S2667145X24000026-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139549479","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Therapeutic drug monitoring of glycopeptide antimicrobials: An overview of liquid chromatography-tandem mass spectrometry methods","authors":"Alessia Cafaro ,&nbsp;Sebastiano Barco ,&nbsp;Federica Pigliasco ,&nbsp;Chiara Russo ,&nbsp;Marcello Mariani ,&nbsp;Alessio Mesini ,&nbsp;Carolina Saffioti ,&nbsp;Elio Castagnola ,&nbsp;Giuliana Cangemi","doi":"10.1016/j.jmsacl.2023.12.003","DOIUrl":"10.1016/j.jmsacl.2023.12.003","url":null,"abstract":"<div><p>Therapeutic drug monitoring (TDM) is a critical clinical tool used to optimize the safety and effectiveness of drugs by measuring their concentration in biological fluids. These fluids are primarily plasma or blood. TDM, together with real-time dosage adjustment, contributes highly to the successful management of glycopeptide antimicrobial therapies. Understanding pharmacokinetic/pharmacodynamic (PK/PD) properties is vital for optimizing antimicrobial therapies, as the efficacy of these therapies depends on both the exposure of the patient to the drug (PK) and pharmacodynamic (PD) parameters such as the in vitro estimated minimum drug concentration that inhibits bacterial growth (MIC). Liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) is widely recognized as the gold standard for measuring small molecules, such as antibiotics. This review provides a comprehensive overview of LC-MS/MS methods available for TDM of glycopeptide antibiotics, including vancomycin, teicoplanin, dalbavancin, oritavancin, and telavancin.</p></div>","PeriodicalId":52406,"journal":{"name":"Journal of Mass Spectrometry and Advances in the Clinical Lab","volume":"31 ","pages":"Pages 33-39"},"PeriodicalIF":2.2,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2667145X23000408/pdfft?md5=4cbaef25bba910aaf200989c874bec54&pid=1-s2.0-S2667145X23000408-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139012527","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Is commutability of a reference material always desirable?","authors":"Michael Vogeser,&nbsp;Katharina Habler","doi":"10.1016/j.jmsacl.2023.12.002","DOIUrl":"10.1016/j.jmsacl.2023.12.002","url":null,"abstract":"","PeriodicalId":52406,"journal":{"name":"Journal of Mass Spectrometry and Advances in the Clinical Lab","volume":"31 ","pages":"Pages 17-18"},"PeriodicalIF":2.2,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2667145X2300041X/pdfft?md5=9a67eab2f5be152a66257f38cbcea7de&pid=1-s2.0-S2667145X2300041X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139016771","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhancing saliva diagnostics: The impact of amylase depletion on MALDI-ToF MS profiles as applied to COVID-19","authors":"Zane LaCasse ,&nbsp;Prajkta Chivte ,&nbsp;Kari Kress ,&nbsp;Venkata Devesh R. Seethi ,&nbsp;Joshua Bland ,&nbsp;Hamed Alhoori ,&nbsp;Shrihari S. Kadkol ,&nbsp;Elizabeth R. Gaillard","doi":"10.1016/j.jmsacl.2024.01.003","DOIUrl":"10.1016/j.jmsacl.2024.01.003","url":null,"abstract":"<div><h3>Introduction</h3><p>Human saliva contains a wealth of proteins that can be monitored for disease diagnosis and progression. Saliva, which is easy to collect, has been extensively studied for the diagnosis of numerous systemic and infectious diseases. However, the presence of amylase, the most abundant protein in saliva, can obscure the detection of low-abundance proteins by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-ToF MS), thus reducing its diagnostic utility.</p></div><div><h3>Objectives</h3><p>In this study, we used a device to deplete salivary amylase from water-gargle samples by affinity adsorption. Following depletion, saliva proteome profiling was performed using MALDI-ToF MS on gargle samples from individuals confirmed to have COVID-19 based on nasopharyngeal (NP) swab reverse transcription quantitative polymerase chain reaction (RT-qPCR).</p></div><div><h3>Results</h3><p>The depletion of amylase led to increased signal intensities of various peaks and the detection of previously unobserved peaks in the MALDI-ToF MS spectra. The overall specificity and sensitivity after amylase depletion were 100% and 85.17%, respectively, for detecting COVID-19.</p></div><div><h3>Conclusion</h3><p>This simple, rapid, and inexpensive technique for depleting salivary amylase can reveal spectral diversity in saliva using MALDI-ToF MS, expose low-abundance proteins, and assist in establishing novel biomarkers for diseases.</p></div>","PeriodicalId":52406,"journal":{"name":"Journal of Mass Spectrometry and Advances in the Clinical Lab","volume":"31 ","pages":"Pages 59-71"},"PeriodicalIF":2.2,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2667145X24000038/pdfft?md5=ef249bcfa77fe37f331dafabc06ba6a6&pid=1-s2.0-S2667145X24000038-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139637213","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development and validation of a liquid chromatography tandem mass spectrometry assay for the analysis of bedaquiline and M2 in breast milk","authors":"Buyisile Mkhize,&nbsp;Richard Court,&nbsp;Sandra Castel,&nbsp;Anton Joubert,&nbsp;Marthinus van der Merwe,&nbsp;Lubbe Wiesner","doi":"10.1016/j.jmsacl.2023.12.001","DOIUrl":"https://doi.org/10.1016/j.jmsacl.2023.12.001","url":null,"abstract":"<div><h3>Objective</h3><p>To develop and validate an assay for the analysis of bedaquiline and its M2 metabolite in human breast milk.</p></div><div><h3>Methods</h3><p>The analytes were extracted using solid phase extraction following protein precipitation. Quantification was performed with liquid chromatography coupled with tandem mass spectrometry. Chromatographic separation was achieved using gradient chromatography on a Poroshell 120 SB-C18 analytical column at 40 °C, with a flow rate of 350 µL/minute and a total run time of eight minutes. An AB Sciex 3000 mass spectrometer with electrospray ionization in the positive mode was used for detection, employing multiple reaction monitoring scan mode. Bedaquiline-d6 and M2-d3-¹³C were used as internal standards.</p></div><div><h3>Results</h3><p>Calibrations curves for bedaquiline and M2 exhibited quadratic (weighted 1/x concentration) regressions over the respective concentration ranges of 0.0780 to 5.00 µg/mL and 0.0312 to 2.00 µg/mL. Inter- and intra-day validation accuracies ranged between 96.7 % and 103.5 % for bedaquiline, and 104.2 % to 106.5 % for M2, with a coefficient of variation below 9.2 % for both compounds.</p></div><div><h3>Conclusion</h3><p>The developed assay demonstrated selectivity and robustness, enabling differentiation between bedaquiline and M2 within the context of endogenous compounds from six separate lots of breast milk samples. Successful application was observed in the analysis of breast milk samples sourced from patients treated for multidrug-resistant tuberculosis within a clinical study setting.</p></div>","PeriodicalId":52406,"journal":{"name":"Journal of Mass Spectrometry and Advances in the Clinical Lab","volume":"31 ","pages":"Pages 8-16"},"PeriodicalIF":2.2,"publicationDate":"2023-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2667145X23000391/pdfft?md5=10be0ce9af33a42047481730cdb66b49&pid=1-s2.0-S2667145X23000391-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138713527","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Use of volumetric absorptive microsampling and parallel reaction monitoring mass spectrometry for tacrolimus blood trough measurements at home in pediatric heart transplant patients","authors":"Junfang Zhao ,&nbsp;Kenneth D.R. Setchell ,&nbsp;Xueheng Zhao ,&nbsp;Stephanie Galandi ,&nbsp;BreAnn N Garr ,&nbsp;Zhiqian Gao ,&nbsp;Clifford Chin ,&nbsp;Shelly Stark ,&nbsp;Paul E. Steele ,&nbsp;Thomas D. Ryan","doi":"10.1016/j.jmsacl.2023.11.004","DOIUrl":"https://doi.org/10.1016/j.jmsacl.2023.11.004","url":null,"abstract":"<div><h3>Background</h3><p>Measurement of trough levels for calcineurin inhibitors by venipuncture sampling is a mainstay of patient management in solid organ transplant recipients but challenging in pediatric patients. Volumetric Absorptive Microsampling (VAMS) is a patient-friendly, minimally invasive sampling technique to accurately collect blood. An assay for measurement of tacrolimus in blood using VAMS, coupled with parallel reaction monitoring (PRM) mass spectrometry, was validated in pediatric heart transplant patients.</p></div><div><h3>Methods</h3><p>Tacrolimus was measured by a newly developed high-resolution PRM assay and compared with low-resolution tandem mass spectrometry (MRM). Dried blood samples were collected from pediatric heart transplant patients (n = 35) using VAMS devices and a satisfaction survey was completed by patients/guardians. Tacrolimus concentrations were compared across whole liquid blood, dried blood spots, and capillary blood, and shipping stability determined.</p></div><div><h3>Results</h3><p>The PRM assay was linear over a range 1–50 ng/mL, similar to MRM but had greater specificity due to reduced background noise. No significant differences in tacrolimus concentrations were observed between VAMS and venous blood. Tacrolimus dried on VAM tips was stable for 14 days and concentrations were unaffected by postal shipping. The variability in two simultaneously collected at-home patient samples was minimal – average concentration difference was 0.12 ± 0.94 ng/mL (<em>p</em> = 0.6) between paired samples.</p></div><div><h3>Conclusion</h3><p>A high resolution PRM mass spectrometry assay was developed for home-based dried blood collections for therapeutic monitoring of tacrolimus. The advantage of PRM was enhanced specificity and the VAMS devices provided a simple and convenient approach to blood sampling at home in pediatric heart transplant patients.</p></div>","PeriodicalId":52406,"journal":{"name":"Journal of Mass Spectrometry and Advances in the Clinical Lab","volume":"31 ","pages":"Pages 1-7"},"PeriodicalIF":2.2,"publicationDate":"2023-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2667145X2300038X/pdfft?md5=84de605382d2d53426e7420203b47793&pid=1-s2.0-S2667145X2300038X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138558643","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exploring Streptococcus pneumoniae capsular typing through MALDI-TOF mass spectrometry and machine-learning algorithms in Argentina: Identifying prevalent NON PCV13 serotypes alongside PCV13 serotypes","authors":"Jonathan Zintgraff ,&nbsp;Florencia Rocca ,&nbsp;Nahuel Sánchez Eluchans ,&nbsp;Lucía Irazu ,&nbsp;Maria Alicia Moscoloni ,&nbsp;Claudia Lara ,&nbsp;Mauricio Santos","doi":"10.1016/j.jmsacl.2023.11.003","DOIUrl":"https://doi.org/10.1016/j.jmsacl.2023.11.003","url":null,"abstract":"&lt;div&gt;&lt;h3&gt;Introduction&lt;/h3&gt;&lt;p&gt;Laboratory surveillance of &lt;em&gt;Streptococcus pneumoniae&lt;/em&gt; serotypes plays a crucial role in effectively implementing vaccines to prevent invasive pneumococcal diseases. The conventional method of serotyping, known as the Quellung reaction, is both time-consuming and expensive. However, the emergence of MALDI-TOF MS technology has revolutionized microbiology laboratories by enabling rapid and cost-effective serotyping based on protein profiles.&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Objectives&lt;/h3&gt;&lt;p&gt;In this study, we aimed to investigate the viability of utilizing MALDI-TOF MS technology as an adjunctive and screening method for capsular typing of &lt;em&gt;Streptococcus pneumoniae&lt;/em&gt;. Our approach involved developing classification models based on MALDI-TOF MS to discern between &lt;em&gt;Streptococcus pneumoniae&lt;/em&gt; strains originating from PCV13 (13-valent pneumococcal conjugate vaccine) and NON PCV13 isolates.&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Methods&lt;/h3&gt;&lt;p&gt;Firstly, we established a comprehensive spectral database comprising isolates of serotypes present in the PCV13 vaccine, along with the top 10 most prevalent NON PCV13 serotypes based on local epidemiological data. This database served as a foundation for developing unsupervised models utilizing MALDI-TOF MS spectra, which enabled us to identify inherent patterns and relationships within the data. Our analysis involved a dataset comprising 215 new isolates collected from nationwide surveillance in Argentina. Our approach involved developing classification models based on MALDI-TOF MS to discern between &lt;em&gt;Streptococcus pneumoniae&lt;/em&gt; strains originating from PCV13 (13-valent pneumococcal conjugate vaccine) and NON PCV13 isolates.&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Results&lt;/h3&gt;&lt;p&gt;Although our findings revealed suboptimal performance in serotype classification, they provide valuable insights into the potential of machine learning algorithms in this context. The sensitivity of the models ranged from 0.41 to 0.46, indicating their ability to detect certain serotypes. The observed specificity consistently remained at 0.60, suggesting a moderate level of accuracy in identifying non-vaccine serotypes. These results highlight the need for further refinement and optimization of the algorithms to enhance their discriminative power and predictive accuracy in serotype identification.&lt;/p&gt;&lt;p&gt;By addressing the limitations identified in this study, such as exploring alternative feature selection techniques or optimizing algorithm parameters, we can unlock the full potential of machine learning in robust and reliable serotype classification of &lt;em&gt;S. pneumoniae&lt;/em&gt;. Our work not only provides a comprehensive evaluation of multiple machine learning models but also emphasizes the importance of considering their strengths and limitations.&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Conclusion&lt;/h3&gt;&lt;p&gt;Overall, our study contributes to the growing body of research on utilizing MALDI-TOF MS and machine learning algorithms for serotype identification purposes.&lt;/p&gt;&lt;/","PeriodicalId":52406,"journal":{"name":"Journal of Mass Spectrometry and Advances in the Clinical Lab","volume":"30 ","pages":"Pages 61-73"},"PeriodicalIF":2.2,"publicationDate":"2023-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2667145X23000366/pdfft?md5=982e9a1f5c8f09914eab8542dde22c4a&pid=1-s2.0-S2667145X23000366-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138453575","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cheaper, faster, simpler trypsin digestion for high-throughput targeted protein quantification","authors":"Christopher M. Shuford,&nbsp;Russell P. Grant","doi":"10.1016/j.jmsacl.2023.11.002","DOIUrl":"https://doi.org/10.1016/j.jmsacl.2023.11.002","url":null,"abstract":"<div><h3>Introduction</h3><p>LC-MS-based methods for protein quantification have a stigma of being relatively expensive and low-throughput. This is partly due to the cost and speed of trypsin digestion, which has primarily focused on advancements in research-based biomarker discovery applications that rely on protein/peptide identifications rather than clinical biomarker quantification. However, there is a need for simple, fast, and reproducibly efficient surrogate peptide recovery in clinical biomarker quantification.</p></div><div><h3>Methods</h3><p>Multiple methodologies were evaluated to enhance tryptic digestion for the analysis of thyroglobulin, a prototypical serum protein biomarker. The main criteria for assessment were the yield and speed of formation of surrogate peptides. Various factors such as different additives, types of trypsin, microwave- and pressure-assisted systems, and enzyme concentration were considered as key variables, in addition to digestion time.</p></div><div><h3>Results</h3><p>It was observed that digestion additives/denaturants had a significant impact on the speed and yield of digestion for each surrogate peptide. Increasing the concentration of trypsin alone was found to accelerate digestions appreciably for most surrogate peptides, without affecting the yield. However, the use of sequencing-grade trypsins and microwave/pressure-assisted systems did not offer significant advantages over the use of 'standard-grade' TPCK-treated trypsin in combination with a conventional incubator, once digestion time and additive had been optimized.</p></div><div><h3>Conclusion</h3><p>We have dispelled the notion that trypsin digestion is inherently slow and expensive for targeted quantification of serum proteins. Additionally, we have established a groundwork for experimentation that can pave the way for the creation of efficient trypsin digestion protocols, aiming to optimize yield, speed, and cost. It is our hope that these advancements will promote the wider incorporation of such assays in clinical laboratories.</p></div>","PeriodicalId":52406,"journal":{"name":"Journal of Mass Spectrometry and Advances in the Clinical Lab","volume":"30 ","pages":"Pages 74-82"},"PeriodicalIF":2.2,"publicationDate":"2023-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2667145X23000378/pdfft?md5=e17d7e7e9bb5f49dff32999123ca1fd3&pid=1-s2.0-S2667145X23000378-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138465856","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}