Bioanalysis最新文献

{"title":"Integrated microfluidic platform for the capture and identification of circulating tumor cells using dielectrophoresis and image recognition.","authors":"Min-Sheng Hung, Sheng-Ze Qiu, Yu-Cheng Wu","doi":"10.1080/17576180.2026.2750760","DOIUrl":"https://doi.org/10.1080/17576180.2026.2750760","url":null,"abstract":"<p><strong>Background: </strong>Circulating tumor cells (CTCs), which detach from primary tumors and enter the peripheral blood, serve as biomarkers for monitoring a patient's status following cancer therapy.</p><p><strong>Methods: </strong>In this study, we developed a microfluidic platform integrating dielectrophoresis and image recognition for the capture and detection of CTCs from blood sample. Experimental parameters included applied electric field strengths ranging from 12.5 to 32.5 × 10⁴ V/m (at 1 MHz) and sample flow rates of 6.25-31.25 mm/min. Fluorescent antibodies were employed to label captured CTCs for real-time detection. Furthermore, a detection software utilizing OpenCV was developed to label CTCs through image processing. The identification performance of the self-developed software was evaluated and compared with machine learning algorithms.</p><p><strong>Results and conclusions: </strong>Results demonstrated that the optimal CTC separation efficiency of 78 ± 2% was achieved at an electric field of 25 × 10⁴ V/m and a flow rate of 18.75 mm/min. While the self-developed software achieved a lower recall than the YOLOv8 model (85 ± 4% vs. 92 ± 2%), its precision matched that of YOLOv8 (90 ± 3% vs. 89 ± 4%).</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"1-10"},"PeriodicalIF":2.1,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149226973","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bioanalytical assays for clinical similarity assessment of biotherapeutics: evolving platforms and regulatory considerations.","authors":"Vikas Chandnani, Sanjay Tiwari, Manoj Bob, Amol Pawar, Suhas Khandave, Supraja Atheriya, Muddukrishna Badamane Sathyanarayana","doi":"10.1080/17576180.2026.2746943","DOIUrl":"https://doi.org/10.1080/17576180.2026.2746943","url":null,"abstract":"<p><p>The rapid expansion of biosimilars following the patent expiry of biologic therapies has increased the need for robust bioanalytical strategies capable of supporting biosimilarity assessment and regulatory decision-making. Within the totality-of-evidence framework, bioanalytical assays play a pivotal role in generating comparative pharmacokinetic (PK), pharmacodynamic (PD), and immunogenicity data that reduce residual uncertainty and demonstrate comparability between a biosimilar candidate and its reference product. The selection of appropriate analytical methodologies is therefore critical to ensuring reliable evaluation of clinical performance, immunogenicity risk, and therapeutic equivalence.This review critically evaluates the application of major bioanalytical platforms used in biosimilar development, including ligand binding assays (LBA), liquid chromatography-tandem mass spectrometry (LC-MS/MS), and hybrid immunoaffinity-LC-MS/MS approaches. Their respective strengths, limitations, and regulatory relevance are discussed in the context of PK quantification, PD biomarker assessment, anti-drug antibody (ADA) detection, and neutralizing antibody (NAb) evaluation. Comparative analysis focuses on key assay performance characteristics such as sensitivity, specificity, selectivity, dynamic range, multiplexing capability, susceptibility to matrix and drug interference, and operational complexity. The contribution of each platform to addressing residual uncertainty during biosimilarity assessment is also examined.Furthermore, the review highlights recent technological advances that have improved assay performance and expanded the analytical toolbox available for complex biologics and emerging biosimilar modalities. A practical decision-making framework is proposed to guide assay selection according to study objectives, biomarker availability, molecular complexity, immunogenicity risk, and the degree of residual uncertainty remaining after analytical similarity assessment. Literature was identified through systematic searches of PubMed and Google Scholar covering publications from 2005 to 2026, with emphasis on recent developments in bioanalytical technologies and evolving regulatory expectations. This review provides a biosimilar-focused perspective on the strategic integration of bioanalytical approaches to support efficient development programs, strengthen the totality of evidence, and facilitate evidence-based regulatory submissions.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"1-14"},"PeriodicalIF":2.1,"publicationDate":"2026-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149219765","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"European Bioanalysis Forum cross-industry perspective on emerging small-molecule modalities: bioanalytical challenges and potential mitigation strategies.","authors":"Luca Ferrari, Cecilia Arfvidsson, Alessandro Greco, Joerg Pausch, Elaine Corcoran, Kristi House, Daniel Mascher, Daniel Schulz-Jander, Sonja Neitzel, Christian Heinlein, Tobias Klaassen, Simon Beuck, Amanda Wilson, Heike Wiese, Michael Blackburn, Stephanie Keane, Rebecca Paterson, Connor Walker, Johanna Paris, Daniel Neddermann, Daniel Villalba, Emmanuel Njumbe Ediage, Philip Timmerman","doi":"10.1080/17576180.2026.2747641","DOIUrl":"https://doi.org/10.1080/17576180.2026.2747641","url":null,"abstract":"<p><p>Emerging small molecule modalities, including oligonucleotides, peptides, lipid nanoparticles, covalent inhibitors and prodrugs, challenge conventional bioanalytical approaches. Recognizing this, the European Bioanalysis Forum convened a cross-industry working group to systematically identify and categorize bioanalytical challenges across seven modality types. Recurring issues include instability, poor recovery, matrix effects and anti-drug antibody interference. Beyond documenting these challenges, the group collected preliminary mitigation strategies based on cross-company experience and scientific discussion. These strategies represent potential approaches rather than formal recommendations. The findings highlight that emerging modalities require early cross-functional assessment and modality-specific analytical strategies, guided by the intended use of the data. This initial categorization provides a foundation for further scientific evaluation, community alignment and regulatory dialogue, supporting the development of science-driven, fit-for-purpose bioanalytical approaches as experience with these modalities evolves.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"1-7"},"PeriodicalIF":2.1,"publicationDate":"2026-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149215113","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optimization of hemolysis index thresholds for biochemical and immunoassays.","authors":"Anne Marie Dupuy, Delphine Tabone, Caroline Coulon, Marion Morena, Jean Paul Cristol","doi":"10.1080/17576180.2026.2744031","DOIUrl":"https://doi.org/10.1080/17576180.2026.2744031","url":null,"abstract":"<p><strong>Background: </strong>Our study aimed at establishing analyte-specific hemolysis-index thresholds on 18 chemistry assays to avoid re-sampling on Atellica© Solution (Siemens Healthcare Diagnostics, Los Angeles, CA, USA).</p><p><strong>Methods: </strong>We focused on the analytes which showed the highest degree of hemolysis interference at one or two baseline concentrations for the following 18 analytes: ALT (alanine aminotransferase), ammonium, AST (aspartate aminotransferase), CK (creatine kinase), creatinine, folates, FSH (follicle stimulating hormone), GGT (γ-glutamyltransferase), Hp (haptoglobin), iron, insulin, LD (lactate dehydrogenase), lipase, potassium, urea, uric acid, vitamins B12, and D. Interference was quantified as a percentage difference from the Hb-free baseline sample. Each calculated bias was compared to the four cutoffs: 10%Δ, analytical change limit, total change limit, or reference change limit.</p><p><strong>Results: </strong>Analytes that did not exceed the predefined allowable-bias criterion for hemolysis included creatinine, FSH, GGT, iron, urea, uric acid, and vitamin B12. Among the affected analytes, the largest positive and negative biases were observed for AST, folates, LD, and potassium. The principal quantitative optimized thresholds for the hemolysis-sensitive analytes were AST, folates, iron, potassium, and LD tests.</p><p><strong>Conclusion: </strong>Our study allowed us to supplement the data provided by the manufacturer and to optimize the thresholds for interference with hemolysis management of hemolyzed samples.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"1-7"},"PeriodicalIF":2.1,"publicationDate":"2026-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149191017","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Applications of enzyme activity assays in clinical development for rare disease therapeutics.","authors":"Catherine Vrentas, Karen J Quadrini, Yan G Ni","doi":"10.1080/17576180.2026.2744071","DOIUrl":"https://doi.org/10.1080/17576180.2026.2744071","url":null,"abstract":"<p><p>Many rare diseases, such as lysosomal storage diseases, are caused by genetically driven enzyme deficiencies. Treatment strategies for these diseases have expanded from substrate reduction and enzyme replacement therapies to more recent gene- and cell-based approaches. As therapeutic modalities for treating rare diseases evolve, bioanalytical assays for measuring enzyme activity have been developed in parallel to support a variety of contexts of use (COUs). In this review, we examine the major contexts in which enzyme activity assays are applied to generate reliable data supporting patient selection, pharmacokinetic, pharmacodynamic, and immunogenicity assessments. Examples from ongoing clinical studies and approved product development are used to illustrate these applications. In particular, we highlight the growing role of enzyme activity assays in pharmacodynamic biomarker assessment and challenges related to preanalytical variability and maintaining assay performance during longitudinal clinical studies. Technical considerations associated with enzyme activity assays for different COUs are also discussed. As therapeutic strategies for rare enzyme deficiency disorders continue to advance, the various COUs that enzyme activity assays support and the corresponding analytical validation approaches may further evolve to inform clinical development and regulatory decision-making.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"1-14"},"PeriodicalIF":2.1,"publicationDate":"2026-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149179330","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A sensitive assay for estimation of DM1 fortified with trastuzumab in human serum utilizing positive LC-MS/MS electrospray ionization and its application to clinical study.","authors":"Ashok Ghoghari, Harilal Patel, Chandrakant Bhatt, Gaurav Koshti, Mukul Jain, Deven Parmar, Ravi Kumar Trivedi","doi":"10.1080/17576180.2026.2746946","DOIUrl":"https://doi.org/10.1080/17576180.2026.2746946","url":null,"abstract":"<p><strong>Aim: </strong>A sensitive LC-MS/MS method was developed and validated for the estimation of DM1 fortified with trastuzumab in human serum.</p><p><strong>Materials and methods: </strong>Quantification was performed in positive mode with protein precipitation extraction using DM4 as an internal standard. Chromatographic separation was achieved using gradient elution and column ACE 5 C18, 100 mm × 4.6 mm. The mobile phase consisted of 0.1% formic acid in 1 mM ammonium formate in water and acetonitrile:methanol (50:50%, v/v) with a total run time of 8.0 min.</p><p><strong>Results: </strong>The calibration curve was linear over the range of 0.5-100 ng/mL. Inter-day and intra-day accuracy and precision, matrix effect, and recovery were well within the acceptance criteria.</p><p><strong>Conclusion: </strong>A method to quantify free DM1 in human serum was fully validated, and the validated assay was successfully applied to determine the clinical pharmacokinetics of DM1 in HER2-positive patients.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"1-11"},"PeriodicalIF":2.1,"publicationDate":"2026-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149179387","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bedside analysis of volatile organic compounds in exhaled human breath as non-invasive diagnostic tool for colon diseases.","authors":"Jakub Pietrzak, Monika Wiśnik-Sawka, Mirosław Maziejuk, Krzysztof Kucharczyk, Adam Przybyłkowski","doi":"10.1080/17576180.2026.2744036","DOIUrl":"https://doi.org/10.1080/17576180.2026.2744036","url":null,"abstract":"<p><strong>Background: </strong>Measurements of volatile organic compounds (VOCs) in exhaled air represent a promising noninvasive biomarker modality for gastrointestinal diseases and has potential to enable point-of-care diagnostics.</p><p><strong>Aim: </strong>To evaluate the feasibility of differential ion mobility spectrometry (DMS) combined with machine learning to distinguish patients with colon diseases from healthy controls based on VOCs analysis in breath.</p><p><strong>Methods: </strong>Breath samples were collected from 50 participants, including patients with inflammatory bowel disease (IBD), diverticulosis and healthy controls. Exhaled VOC profiles were acquired using a bedside differential mobility spectrometry (DMS) platform and subsequently analyzed with machine learning models.</p><p><strong>Results: </strong>Based on machine-learning analysis there was moderate discrimination between patients with colon disease and healthy controls (best ROC-AUC 0.784). In exploratory disease-specific analyses the highest discriminative performance for ulcerative colitis (ROC-AUC 0.862) has been noted, whereas discrimination between Crohn's disease and diverticular disease was less accurate. In patient-level permutation testing it has been confirmed that the observed diagnostic performance was unlikely to have occurred by chance (permutation <i>p</i> = 0.005).</p><p><strong>Conclusion: </strong>DMS-based breath analysis combined with machine learning appears promising noninvasive diagnostic approach in patients with colon diseases.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"1-8"},"PeriodicalIF":2.1,"publicationDate":"2026-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149168727","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development and pre-qualification of a facile and ultra-sensitive LRRK2 LC/MS assay using high-affinity unmodified immobilization for immunocapture: meet bioanalytical challenges and search practical solutions.","authors":"Yu Gus Hui, Jian Wu, Kumar Shah, Roberto C Mendez, Xueqian Su, Fang Wang, Michael Borrome, M Shane Woolf, Moucun Yuan, William Mylott","doi":"10.1080/17576180.2026.2744032","DOIUrl":"https://doi.org/10.1080/17576180.2026.2744032","url":null,"abstract":"<p><strong>Background: </strong>Quantifying low-abundance LRRK2 in CSF is analytically challenging but remains critical for Parkinson's disease biomarker development.</p><p><strong>Methods: </strong>An immunocapture LC-MS assay using high-affinity unmodified antibody immobilization on Protein G beads coupled with a Vanquish Neo-Stellar nano-LC-MS platform was developed and pre-qualified.</p><p><strong>Results: </strong>In 250 µL CSF, the assay achieved an LLOQ of 2 pg/mL, reduced antibody consumption (0.8 µg/sample), robust surrogate-to-matrix parallelism, and acceptable accuracy and precision under fit-for-purpose criteria.</p><p><strong>Conclusions: </strong>This practical, ultra-sensitive workflow overcomes key limitations of prior SISCAPA-based methods and is suitable for high-throughput translational and clinical LRRK2 biomarker studies. This flexible framework is extendable to other low-abundance protein biomarkers requiring rugged, automated, nano-LC mass spectrometric quantification in complex biological matrices.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"1-15"},"PeriodicalIF":2.1,"publicationDate":"2026-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149126537","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"From urine to serum: blood-based matrices reshape the future of anti-doping testing.","authors":"Tobias Langer, Tiia Kuuranne, Nicolas Leuenberger","doi":"10.1080/17576180.2026.2736404","DOIUrl":"10.1080/17576180.2026.2736404","url":null,"abstract":"<p><p>For more than five decades, urine has been the cornerstone biological matrix for anti-doping testing because of its noninvasive collection, broad analytical applicability, and favorable detection windows. However, evolving doping practices, including microdosing, increased use of biological medicines, and substances with limited urinary excretion, have exposed limitations of a urine-centered approach. At the same time, advances in liquid chromatography-tandem mass spectrometry (LC-MS/MS), high-resolution mass spectrometry (HRMS), and microsampling technologies have strengthened the potential of blood-based testing. Serum and dried blood spots (DBS) provide direct access to circulating analytes and physiological responses, offering complementary information to urine. These matrices have demonstrated particular value for detecting erythropoiesis-stimulating agents (ESAs), steroid esters, peptide hormones, and biomarkers incorporated into the Athlete Biological Passport (ABP). Emerging capillary blood collection devices further simplify sample collection and may facilitate wider implementation of blood-based anti-doping strategies. This review discusses the expanding role of serum, DBS, and capillary blood collection technologies in anti-doping analysis. Using ESA detection as an illustrative example, we examine how matrix selection affects detection capability and analytical performance. We also highlight applications in steroid monitoring, biological profiling, and future omics-based approaches, supporting the integration of blood-derived matrices into complementary multi-matrix anti-doping testing strategies.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"1107-1116"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13644600/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149011751","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":"Recent developments in microsampling for oligonucleotides and biologics bioanalysis.","authors":"Guilherme Guimaraes, Long Yuan","doi":"10.1080/17576180.2026.2739128","DOIUrl":"10.1080/17576180.2026.2739128","url":null,"abstract":"<p><p>The expanding landscape of therapeutic modalities, including monoclonal antibodies and oligonucleotide-based drugs, has created a need for sensitive and efficient bioanalytical strategies. Traditional blood sampling approaches pose significant limitations, including volume constraints, patient burden, and increased study complexity. Microsampling provides a transformative alternative, enabling low-volume (<100 µL), minimally invasive sample collection while supporting serial sampling and improved translational study design. This review highlights recent advances in microsampling techniques, including dried blood spots (DBS), volumetric absorptive microsampling (VAMS), and capillary microsampling (CMS), with focus on applications to protein therapeutics and oligonucleotides. Selected biomarker and protein/proteomics applications are also discussed where they provide practical insight into microsampling implementation for larger modalities. Key challenges, including hematocrit effects, recovery, matrix-dependent bias, stability, collection-site effects, and sensitivity limitations, are discussed alongside innovations in hybridization-based extraction, ligand-binding assays, and LC-MS workflows that enable reliable quantitation in microliter-scale samples. Despite these advances, broader implementation in regulated settings remains limited by regulatory considerations and the need to demonstrate concordance with conventional sampling. Ongoing efforts to harmonize best practices and improve analytical robustness are expected to accelerate adoption. Overall, microsampling offers opportunities to reduce burden, enhance efficiency, and expand access to high-quality PK data across emerging therapeutic modalities.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":"18 12-13","pages":"1183-1196"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13644594/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149103574","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}