Aravapally PSN, Chandrasekar N, Verma A
… +1 more, Shah RP
Bioanalysis
· 2025 Apr · PMID 40183176
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Oxidative stress (OS) is an emerging research area in clinical and biological sciences due to its association with various diseases and physiological processes. OS occurs when there is an imbalance between the production...Oxidative stress (OS) is an emerging research area in clinical and biological sciences due to its association with various diseases and physiological processes. OS occurs when there is an imbalance between the production of reactive oxygen species (ROS) and the body's ability to neutralize or repair the damage caused. Chronic oxidative stress is linked to diseases like diabetes, cardiovascular diseases, cancer, and neurodegenerative disorders. Accurate monitoring of OS is crucial for diagnosing diseases, evaluating disease progression, and predicting clinical results. Despite challenges in measuring free radicals due to their short half-life and low concentrations, it can be indirectly assessed through biomarkers like lipid peroxidation, DNA damage, and protein oxidation. The most effective analytical techniques for assessing OS biomarkers in various biological fluids were developed. Furthermore, an in-depth exploration of these various analytical methodologies, underscoring their sensitivity, specificity, and reliability in detecting low concentrations of biomarkers across complex matrices is necessary. A comprehensive literature search was conducted using databases such as Google Scholar, PubMed and Reaxys to identify relevant studies on OS biomarkers. This review explores the evolution of these techniques, highlighting advancements in sample preparation procedures and the specifications of each technique, offering a thorough evaluation of biomarker analysis.
Bioanalysis
· 2025 Mar · PMID 40153274
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Immunosuppressive pharmacotherapy after solid organ transplantation (SOT) requires therapeutic drug monitoring (TDM) for therapy individualization. The venous whole blood is still considered as routine matrix for monitor...Immunosuppressive pharmacotherapy after solid organ transplantation (SOT) requires therapeutic drug monitoring (TDM) for therapy individualization. The venous whole blood is still considered as routine matrix for monitoring immunosuppressive drug concentration. On the other hand, as an alternative, capillary blood collected using noninvasive sampling is convergent with a patient-centric approach. Despite their disadvantages regarding sample homogeneity and the hematocrit effect, well-known dried blood spot techniques have shown promising results. Volumetric absorptive microsampling (VAMS) and quantitative dried blood spot (qDBS) have successfully eliminated these unfavorable biased elements. Microsampling can be used in transplant recipients' care, mainly due to long-term therapy under control drug concentrations and the long distance between the place of the patient's residence and the diagnostic laboratory in the transplant center. The study aimed to discuss the clinical consequences of implementing microsampling techniques for TDM of immunosuppressants. Additionally, we have discussed the 'hot topics' in microsampling: home-based self-sampling, adherence to therapy monitoring, and drug concentration conversion to estimated traditional matrices. Finally, based on our experience and current practice, we propose best practices for microsampling implementation from bench to bedside. Microsampling techniques can potentially revolutionise immunosuppressive pharmacotherapy by enabling patient-centric individualisation in various subpopulations, significantly improving post-transplant care.
Ahmad S, Gärtner F, Penford R
… +22 more, Wolter L, Belle S, Bushby A, Carneiro Da Cunha M, Dan B, Di Ianni A, Dumolyn C, Giordano MS, Hanckmann E, Maher J, Milandri E, Pineault K, Roberts A, Reille-Seroussi M, Russo S, Spendal M, Szuster C, Vandenbroucke K, Vollmer A, Vogt F, Wellenberg S, Timmerman P
Bioanalysis
· 2025 Apr · PMID 40131210
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Since 2014, the European Bioanalysis Forum has organized the Young Scientist Symposium, providing early-career scientists with unique development opportunities. By fostering a peer community, this initiative has successf...Since 2014, the European Bioanalysis Forum has organized the Young Scientist Symposium, providing early-career scientists with unique development opportunities. By fostering a peer community, this initiative has successfully lowered barriers to engagement in cross-company, pre-competitive interactions. Since 2018, the symposium has included the Science Café roundtable, further reinforcing the European Bioanalysis Forum's commitment to supporting young scientists beyond their scientific expertise by promoting broader professional development.This manuscript summarizes discussions from the 10th Young Scientist Symposium, held in Hasselt, Belgium, from May 15-17, 2024. The symposium featured presentations on new technologies, biomarker assays, troubleshooting, and microsampling. Additionally, it provides insights from the Science Café discussions, which focused on sustainability in the bioanalytical laboratory.
Jain R, Jain B, Al-Khateeb LA
… +4 more, Alharthi S, Ghoneim MM, AbdElrahman M, Alanazi AS
Bioanalysis
· 2025 Apr · PMID 40126928
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Bioanalytical laboratories face significant challenges in sample preparation due to the complexity of biological matrices and the low concentrations of target analytes. This review focuses on advances in green sample pre...Bioanalytical laboratories face significant challenges in sample preparation due to the complexity of biological matrices and the low concentrations of target analytes. This review focuses on advances in green sample preparation (GSP) techniques tailored to meet these challenges while promoting sustainability. Innovations in sorbents, including metal-organic frameworks (MOFs), magnetic nanoparticles (MNPs), sol-gel-based materials, molecularly imprinted polymers (MIPs), carbon-based materials, and natural sorbents like cellulose and kapok fiber, have enhanced extraction efficiency and selectivity. Green solvents such as deep eutectic solvents (DES), ionic liquids (ILs), supramolecular solvents (SUPRAs), and switchable hydrophilicity solvents (SHSs) further reduce environmental impact by minimizing toxic solvent use. This review highlights their use in drug analysis, emphasizing their roles in enhancing extraction efficiency, selectivity, and environmental sustainability. Recent applications demonstrate the integration of these sorbents and solvents into bioanalytical workflows, significantly improving analytical performance while adhering to Green Analytical Chemistry (GAC) principles. It is anticipated that this comprehensive review will aid scholars in the formulation of selective, rapid, environmentally friendly, straightforward, sensitive, and precise analytical methodologies for bioanalysis, thereby promoting innovation and sustainability within drug analysis protocols.
Mounika P, Shelby M, K S C
… +2 more, Kumar HY, Gurupadayya B
Bioanalysis
· 2025 Apr · PMID 40126089
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AIM: The main purpose of the study is to establish Bioanalytical method development for enantiomeric divergence and Stereoselective Pharmacokinetic activity of 1-(4-bromophenyl)-6,7-dimethoxy-3,4-dihydroisoquinoline-2(1H...AIM: The main purpose of the study is to establish Bioanalytical method development for enantiomeric divergence and Stereoselective Pharmacokinetic activity of 1-(4-bromophenyl)-6,7-dimethoxy-3,4-dihydroisoquinoline-2(1H)-sulfonamide(B06). BACKGROUND: Isoquinoline derivatives have various activities like anticancer, anti-convulsant etc. The proposed project is mainly focused on anti-cancer activity of the synthesized B-06 compound as it shown very good cytotoxicity activity on MBA-MD-231 and MCF-7 cell lines by using SRB assay. METHOD: The specified project was completed with HPLC and an amylose chiral column. The bioanalytical approach was developed and tested using Wister Albino rats in compliance with USFDA Guidelines and expanded to include pharmacokinetic activities. RESULT: Data that had previously been published was used to design the B06 compound. RT of 9.578 and 7.076 minutes were observed for (R) &; (S) B06 Compound. Within-run and between-run precision for S-enantiomer varied from 0.28 to 6.078%, while R-enantiomer was found to range from 0.34 to 6.08%. Recovery rates ranged from 80.06% to 92.60% for both enantiomers. Pharmacokinetic investigations were developed and validated by using PKSolver software with Microsoft Excel. CONCULSION: We successfully established optimized bioanalytical method for pharmacokinetic study were both the enantiomers were properly separated. In pharmacokinetic activity we found that the enantiomers are non-stereoselective having same absorption rate.
Zhang XT, Blacutt J, Lloyd T
… +5 more, Mencer M, Pratt V, Kotha J, Sheeran L, Adcock S
Bioanalysis
· 2025 Mar · PMID 40118816
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Pharmacogenomics (PGx) is transforming therapeutic development by providing insights into how genetic variations influence drug response, safety, and efficacy. This review provides a structured analysis of PGx in clinica...Pharmacogenomics (PGx) is transforming therapeutic development by providing insights into how genetic variations influence drug response, safety, and efficacy. This review provides a structured analysis of PGx in clinical research, beginning with an overview of key genes involved in drug metabolism, transport, and targets. Following this, it examines strategies for identifying PGx-relevant genes, including phenotype-driven, hypothesis-driven, population-focused, and clinical-driven approaches. Technical platforms such as PCR, MassARRAY, and next-generation sequencing are analyzed for their suitability in PGx studies. The discussion then shifts to assay validation processes, covering both analytical and clinical validation, to ensure data reliability in clinical trials. Finally, regulatory expectations for PGx in clinical trials are discussed, focusing on key requirements across all phases of drug development. This review aims to provide a clear and practical framework for integrating PGx into clinical research to enhance drug safety, efficacy, and personalized medicine.
Bioanalysis
· 2025 Mar · PMID 40118813
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BACKGROUND: Pocapavir is an antiviral agent that has shown in vitro potency against poliovirus. It necessitates investigation into its efficacy and safety profiles. Therefore, the development of a sensitive and reliabl...BACKGROUND: Pocapavir is an antiviral agent that has shown in vitro potency against poliovirus. It necessitates investigation into its efficacy and safety profiles. Therefore, the development of a sensitive and reliable bioanalytical method to quantify pocapavir drug levels and characterize its pharmacokinetic (PK) profiles is imperative. RESEARCH DESIGN AND METHODS: Given pacapavir's low ionization efficiency, the utilization of atmospheric pressure chemical ionization (APCI) is employed to enhance the assay sensitivity. The developed method is validated and subsequently implemented in a mouse PK study. RESULTS: As the first reported bioanalytical method for pocapavir, the method is developed and validated through a full method validation in accordance with the principles of Good Laboratory Practice (GLP), with all evaluated parameters meeting the stipulated acceptance criteria. CONCLUSIONS: The validated bioanalytical method is suitably equipped to quantify pocapavir while bolstering preclinical and potential applications for first-in-human (FIH) PK characterizations.
Bioanalysis
· 2025 Apr · PMID 40114295
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PROTACs are reshaping drug discovery by enabling targeted protein degradation, overcoming the limitations of traditional inhibitors, and addressing previously "undruggable" proteins. The present perspective explores adva...PROTACs are reshaping drug discovery by enabling targeted protein degradation, overcoming the limitations of traditional inhibitors, and addressing previously "undruggable" proteins. The present perspective explores advancements in PROTAC molecular design, focusing on ligand discovery, E3 ligase recruitment, and ternary complex optimization. Integrating AI-driven modeling, FBDD, and SBDD accelerates PROTAC development. In contrast, emerging innovations, such as PHOTACs, hypoxia-responsive systems, and Ab-PROTACs, enhance precision and reduce systemic toxicity. Clinical successes, including ARV-110 for castration-resistant prostate cancer and ARV-471 for breast cancer, exemplify their ability to overcome resistance and provide durable effects. PROTACs are expanding into neurodegenerative diseases and rare conditions, highlighting their versatility. By addressing challenges in pharmacokinetics, safety, and scalability, PROTACs are poised to revolutionize precision medicine. This article presents a forward-looking perspective on conquering the molecular design and drugability of PROTACs, paving the path for transformative therapies.
Timmerman P, White S, Adcock N
… +14 more, Arfvidsson C, Barfield M, Cowan K, Ferrari L, Golob M, Goodwin L, Hughes R, Ivanova T, Laurén A, McDougall S, Nelson R, van de Merbel N, Verhaeghe T, Wright M
Bioanalysis
· 2025 Mar · PMID 39985138
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The release of the ICH M12 Guideline on Drug Interaction Studies has reignited discussions around assay validation requirements for assays such as plasma protein-binding studies. Even though the ICH M12 does not directl...The release of the ICH M12 Guideline on Drug Interaction Studies has reignited discussions around assay validation requirements for assays such as plasma protein-binding studies. Even though the ICH M12 does not directly reference the ICH M10 Guideline on Bioanalytical Method Validation and Sample Analysis, its release prompted further discussions on assay validation requirements for these studies during the 17th European Bioanalysis Forum Open Symposium held in Barcelona, Spain, from 19 to 21 November 2024, where we advocated for a Context-of-Use driven approach over rigid adherence to ICH M10 standards. Context-of-Use driven validation ensures assays are tailored to the specific scientific and regulatory needs, optimizing resource allocation and innovation in drug development. This short opinion paper explores the potential and undesired implications of ICH M12 on bioanalytical validation practices, highlights the distinction between exploratory assays and assays having a clinical impact, and underscores the necessity for tailored validation strategies.
Sun W, Mou S, Huntington C
… +14 more, Killick H, Scott IC, Kelly A, Gavala M, Larsson J, Vakkalanka MD, Alexis NE, Wiley W, Wheeler A, Shah K, Yuan M, Mylott WR, Contrepois K, Rosenbaum AI
Bioanalysis
· 2025 Feb · PMID 39976267
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AIM: Airway mucins in sputum are promising respiratory disease biomarkers, despite posing substantial analytical challenges due to their physicochemical properties and rare and heterogenous nature of the matrix. We aimed...AIM: Airway mucins in sputum are promising respiratory disease biomarkers, despite posing substantial analytical challenges due to their physicochemical properties and rare and heterogenous nature of the matrix. We aimed to identify a suitable sputum collection and processing method, and qualify a bioanalytical method for MUC5AC and MUC5B quantification in clinical samples. METHOD: Mucins were quantified in induced and spontaneous sputum collected from the same COPD patients, following various sample processing procedures. LC-MS/MS method used truncated recombinant mucins as surrogate analytes in surrogate matrix. RESULTS: Frozen spontaneous sputum was found to be a suitable and convenient matrix for mucin quantification and fit-for-purpose method qualification was performed. CONCLUSION: Our methodology provides accurate and reliable MUC5AC and MUC5B quantification and facilitates multi-site clinical sputum collection.
Bioanalysis
· 2025 Mar · PMID 39932032
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Singlicate anti-drug antibody (ADA) analysis is gaining recognition as a promising approach to optimize immunogenicity assessment. It offers significant benefits in terms of cost-effectiveness and aligns with patient-cen...Singlicate anti-drug antibody (ADA) analysis is gaining recognition as a promising approach to optimize immunogenicity assessment. It offers significant benefits in terms of cost-effectiveness and aligns with patient-centric principles. However, its successful implementation requires a thorough understanding of its impacts on assay performance. In this perspective, we review recent case studies comparing singlicate and duplicate formats, focusing on cut point factors and sample result agreement, and seek to discuss how singlicate influences assay cut points and explore considerations for low positive control criteria under singlicate conditions. Furthermore, we propose a strategy for implementing singlicate analysis in both new and existing ADA assays, hoping to contribute to ongoing discussions in this area.
Lian K, Lei H, Yang X
… +3 more, Yan J, Yu Y, Li H
Bioanalysis
· 2025 Feb · PMID 39924897
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AIM: The purpose of this study is to establish a light-initiated chemiluminescence assay (LiCA) for the quantitative analysis of pollen-specific IgE (sIgE) antibodies. METHODS: The best chemibeads coupling method in det...AIM: The purpose of this study is to establish a light-initiated chemiluminescence assay (LiCA) for the quantitative analysis of pollen-specific IgE (sIgE) antibodies. METHODS: The best chemibeads coupling method in detecting pollen - sIgE was selected. The working concentrations of the antigen-antibody and the reaction buffer were optimized as components of the reaction system. Then the assay performance was evaluated and the results of LiCA were compared with ImmunoCAP methods. RESULTS: In the range of 0.23 kU/L to 100.51 kU/L, LiCA demonstrated good linearity. The coefficients of variation for repeatability and intermediate precision ranged from 5.96% to 8.58% and 7.53% to 11.25%, respectively. The limit of blank (LoB), limit of detection (LoD), and limit of quantitation (LoQ) were determined to be 0.044 kU/L, 0.086 kU/L, and 0.11 kU/L, respectively. Furthermore, LiCA exhibited a statistically significant correlation with ImmunoCAP ( = 0.990). CONCLUSION: The established LiCA-based quantitative detection method for pollen-slgE has good analytical performance and potential clinical application prospects.
Thomas A, Walpurgis K, Naumann N
… +2 more, Piper T, Thevis M
Bioanalysis
· 2025 Mar · PMID 39916648
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The analytical and technological approaches employed in doping analysis are constantly reviewed and updated to allow for keeping pace with progresses in pharmaceutical and medicinal research and the therein inherent opti...The analytical and technological approaches employed in doping analysis are constantly reviewed and updated to allow for keeping pace with progresses in pharmaceutical and medicinal research and the therein inherent options of misuse as performance enhancing drugs or methods. Enormous changes, improvements, and developments have been achieved in recent years, particularly, but not exclusively, in the bioanalytical sector. Several of these new strategies are examined systematically in this review using examples from the World Anti-Doping Agency (WADA) list of banned substances and methods. The review includes, among others, the application of sophisticated new models mimicking multi compartment models, investigation into new long-term metabolites for anabolic agents, the impact of a distinct gene mutation on the analysis of erythropoietin, studies on the development of new therapeutic protein-based drugs with myostatin inhibiting properties, methods applying the new molecular biological section used to uncover gene doping, and finally new approaches uncovering the prohibited use of autologous blood transfusion. All of these challenges and investigations support the ongoing progress in modern doping controls in the future and will help to fill the gap between the advance of cheating athletes and sport drug testing.
Wessels AMA, Junier LAT, Touw D
… +1 more, Stevens J
Bioanalysis
· 2025 Mar · PMID 39916568
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BACKGROUND: Plasma clearance of iohexol is used to measure glomerular filtration rate, for which a UHPLC-MS/MS analytical method was previously developed. In real-world conditions, samples may be thawed on arrival and sa...BACKGROUND: Plasma clearance of iohexol is used to measure glomerular filtration rate, for which a UHPLC-MS/MS analytical method was previously developed. In real-world conditions, samples may be thawed on arrival and sampled in unvalidated matrices, prompting the need for an improved validation. We aim to improve the method for iohexol determination in plasma with enhanced stability testing, optimized calibration curves, and partial validation in additional matrices. METHODS: Stability testing was conducted up to 9 weeks at room temperature, 4°C, 37°C, and at 37°C with daily two-hour exposure to 55°C. Quintuplicate QC samples were analyzed on 3 days, comparing results from eight-point and two-point calibration curves. Matrix comparison was performed on quintuplicate QC samples in serum, heparin plasma, urine, EDTA whole blood, and heparin whole blood. RESULTS: The method improvements were all compliant with the requirements for bioanalytical methods issued by the US FDA and European Medicines Agency. CONCLUSION: Human EDTA plasma samples can be stored up to 9 weeks at room temperature, 4°C, 37°C, and 37°C with 55°C daily temperature spikes. The samples can be analyzed using a two-point calibration curve and are partial validated for serum-, heparin plasma-, urine-, EDTA whole blood-, and lithium whole blood iohexol samples.
Bioanalysis
· 2025 Mar · PMID 39902785
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Biological accelerator mass spectrometry (AMS) provides ultrasensitive carbon-14 isotopic analysis enabling a deeper understanding of human health concerns by enabling quantification of pharmacokinetics and other molecul...Biological accelerator mass spectrometry (AMS) provides ultrasensitive carbon-14 isotopic analysis enabling a deeper understanding of human health concerns by enabling quantification of pharmacokinetics and other molecular endpoints directly in humans. It enables environmentally and human relevant studies of metabolic pathways through the use of very low concentrations of labeled metabolic substrates in cells and organisms. Here, we discuss why AMS is an important tool for the biosciences, the development and evolution of biological AMS at Livermore and discuss technical refinements that will improve the efficiency of operation for the measurement of ultra-trace levels of C, which, long term, will enable greater ease of use and sample throughput.
Bioanalysis
· 2025 Feb · PMID 39902531
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CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) has transformed molecular biology through its precise gene-editing capabilities. Beyond its initial applications in genetic modification, CRISPR has emer...CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) has transformed molecular biology through its precise gene-editing capabilities. Beyond its initial applications in genetic modification, CRISPR has emerged as a powerful tool in diagnostics and biosensing. This review explores its transition from genome editing to innovative detection methods, including nucleic acid identification, single nucleotide polymorphism (SNP) analysis, and protein sensing. Advanced technologies such as SHERLOCK and DETECTR demonstrate CRISPR's potential for point-of-care diagnostics, enabling rapid and highly sensitive detection. The integration of chemical modifications, CRISPR-Chip technology, and enzymatic systems like Cas12a and Cas13a enhances signal amplification and detection efficiency. These advancements promise decentralized, real-time diagnostic solutions with significant implications for global healthcare. Furthermore, the fusion of CRISPR with artificial intelligence and digital health platforms is paving the way for more accessible, cost-effective, and scalable diagnostic approaches, ultimately revolutionizing precision medicine.
Bioanalysis
· 2025 Mar · PMID 39901692
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Immunogenicity knowledge and the analytical capabilities to characterize molecules have evolved within the last decade. This creates opportunities in Biosimilar development by applying new strategies to demonstrate simil...Immunogenicity knowledge and the analytical capabilities to characterize molecules have evolved within the last decade. This creates opportunities in Biosimilar development by applying new strategies to demonstrate similarity between a proposed Biosimilar to its Reference.Within Immunogenicity Risk Assessment for Biosimilars, in silico and in vitro immunogenicity assessment tools are being evaluated for their utility in Biosimilar development. An ISI including an Immunogenicity Risk Assessment is recommended for Biosimilars within the dossier of licensing applications to facilitate the review by Health Authorities and to explain the immunogenicity in conjunction with analytical and clinical data. Operational aspects should also be considered to refine immunogenicity testing of Biosimilars, e.g. S/N ratio and singlicate sample analysis.
Zhang G, Yan S, Liu Y
… +3 more, Du Z, Min Q, Qin S
Bioanalysis
· 2025 Feb · PMID 39895280
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Undruggable targets account for roughly 85% of human disease-related targets and represent a category of therapeutic targets that are difficult to tackle with traditional methods, but their considerable clinical importan...Undruggable targets account for roughly 85% of human disease-related targets and represent a category of therapeutic targets that are difficult to tackle with traditional methods, but their considerable clinical importance. These targets are generally defined by planar functional interfaces and the absence of efficient ligand-binding pockets, making them unattainable for conventional pharmaceutical strategies. The advent of oligonucleotide-based proteolysis-targeting chimeras (PROTACs) has instilled renewed optimism in addressing these challenges. These PROTACs facilitate the targeted degradation of undruggable entities, including transcription factors (TFs) and RNA-binding proteins (RBPs), via proteasome-dependent mechanisms, thereby presenting novel therapeutic approaches for diseases linked to these targets. This review offers an in-depth examination of recent progress in the integration of PROTAC technology with oligonucleotides to target traditionally undruggable proteins, emphasizing the design principles and mechanisms of action of these innovative PROTACs.