Bioanalysis最新文献

Background: Despite recent advances in capillary blood collection technologies for implementation in a patient centric healthcare world, a direct comparison of various technologies has yet to be performed.

Research design and methods: A cross-sectional comparative study was performed by recruiting healthy subjects (n = 41) to participate in a collection comparison study consisting of four upper arm capillary collection technologies, a traditional fingerstick, and a venous collection. User experience (e.g. pain, preference), collection performance (e.g. sample volume and quality, collection time), and clinical accuracy with respect to venipuncture results were compared across technologies. In addition, healing was assessed by reaching out to subjects at two different time points following collection.

Results: User experience and collection performance results varied across the different capillary collection technologies and a single collection technology did not stand out as superior among those investigated. As correlative results were similar across all technologies, selection of a technology may be based on the intended use population and/or other factors.

Conclusions: Although there are still advancements to be made as well as additional studies to further evaluate the analyte equivalency of capillary and venous specimens, these technologies present a promising patient-centric option to provide clinical results and improve patient care.

Mass spectrometry imaging (MSI) enables the visualization of hundreds to thousands of analytes in biological tissues. MSI is also capable of mapping time-dependent processes that, combined with these static metabolite profiles, provides a clearer picture of the molecular underpinnings of tissue function. This perspective is organized into sections demonstrating how MSI-based methods can provide unique functional data on systems ranging from single step enzyme-catalyzed transformations to complex metabolic network activities and cellular dynamics. This multisystem capability can be exploited to provide detailed descriptions of the molecular mechanisms contributing to tissue function. An aspect missing in many studies are corresponding maps of tissue microenvironments including oxygenation and pH which influence functional activities. Some progress has been made to map hypoxic and acidic tissue using MSI methods, but further development is needed. This includes pairing in vivo MSI functional studies to in vitro models. Additionally, integrating the capabilities of other imaging methods, such as magnetic resonance and vibrational spectroscopy, that are proven to detect tissue microenvironments, with dynamic MSI methods offer a route to match environment with functional activities. The combination of static molecular profiles, metabolic and cellular dynamics, and environmental mapping will provide the most detailed understanding of tissue function.

Collaboration between pharmaceutical companies and contract research organizations is central to drug development, yet projects often stumble on well-known weaknesses: communication that does not adapt to the phase of work, expectations that remain unclear, partnerships reduced to transactions, and functions working in silos. Within the European Bioanalysis Forum, two dedicated teams examined these issues in depth, focusing on communication and on building sustainable partnerships. Their reflections were calibrated through dialogue with the wider community during a Focus Workshop, and sharpened by the perspective of the Forum's leadership. This paper brings forward a number of principles that, in the view of the teams and delegates, are essential to more reliable collaboration. They are not an exhaustive list, but a structured starting point for reflection. Reliable collaboration enables science to move forward efficiently and, ultimately, to benefit the patient.

The 19^th Workshop on Recent Issues in Bioanalysis (19^th WRIB) took place in New Orleans, LA, USA on April 7-11, 2025. Over 1200 professionals representing pharma/biotech companies, CROs, and multiple regulatory agencies convened to actively discuss the most current topics of interest in bioanalysis. The 19^th WRIB included 3 Main Workshops and 7 Specialized Workshops that together spanned 1 week to allow an exhaustive and thorough coverage of all major issues in bioanalysis of biomarkers, immunogenicity, gene therapy, cell therapy and vaccines.Moreover, in-depth workshops on "Implementation Practice for the Newest ELN/LIMS Systems" and on "Vaccine Cell-Based/Functional & Molecular Assays as part of the harmonization of vaccine clinical assays global initiative" were the special features of the 19^th edition.As in previous years, WRIB continued to gather a wide diversity of international, industry opinion leaders and Regulatory Agency experts working on both small and large molecules as well as gene, cell therapies and vaccines to facilitate sharing and discussions focused on improving quality, increasing regulatory compliance, and achieving scientific excellence on bioanalytical issues.This 2025 White Paper encompasses recommendations emerging from the extensive discussions held during the workshop and is aimed to provide the bioanalytical community with key information and practical solutions on topics and issues addressed, in an effort to enable advances in scientific excellence, improved quality and better regulatory compliance. Due to its length, the 2025 edition of this comprehensive White Paper has been divided into three parts for editorial reasons.This publication (Part 3) covers in the Part 3A the recommendations on Gene Therapy, Cell therapy, Vaccines and Biotherapeutics Immunogenicity and in Part 3B the Regulatory Inputs on these topics. Part 1 (Mass Spectrometry Assays and Regulated Bioanalysis/BMV) and Part 2 (Biomarkers/BAV, IVD/CDx, LBA and Cell-Based Assays) are published in volume 18 of Bioanalysis, issues 3 and 2, respectively.

Human routine doping controls rely to a considerable extent on urine samples, and test methods have been optimized for decades to allow for comprehensive analyses with adequate analytical sensitivities. However, attempts of sample manipulation, presumably in order to escape an adverse analytical finding, were reported to include adulteration as well as substitution, which has revived the debate as to when a sample submitted as urine specimen for sports drug testing purposes is, indeed, to be considered as 'urine.' In consideration of selected case reports as well as analytical parameters routinely acquired with doping control urine samples, options concerning guidance in that context are discussed, as detecting and reporting urine substitution and urine adulteration require laboratory approaches that might differ considerably from protocols applied for, e.g. the confirmation of the presence of a prohibited substance. In particular, commonly monitored markers characterizing urinary specifics such as pH, specific gravity, and the presence of a human urinary steroid profile and 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR) in established reference ranges could assist in defining acceptance criteria for urine samples and, thereby, guidelines when a specimen does not qualify as human urine.

Introduction: Routine conjugation protocols are typically used by bioanalytical laboratories for production of their assay critical reagents. Novel molecules can pose unique challenges to the production of high-quality conjugated critical reagents required for clinical bioanalytical assays. Using routine conjugation protocols, we observed gross instability of conjugated-drug surrogate material for use in antidrug antibody (ADA) assays in clinical autologous Chimeric Antigen Receptor (CAR)-T cell programs, thus halting assay development.

Methods: We highlight our approach in developing process conditions for CAR-T supporting conjugated critical reagents, where recombinant CAR scFv-hFc is the surrogate drug material to be conjugated. We show that when routine platform process approaches are not appropriate to produce conjugated novel molecule critical reagents, in-silico modeling can be used to determine conditions imparting repeatable generation of stable reagents. Implementing this modeling is advantageous as it decreases laborious efforts to develop stable novel critical reagent production methodologies.

Conclusion: As a result of these studies herein, optimal stable conjugated critical reagents were produced which enabled successful development and validation of clinical CAR-T ADA assay. This work provides a framework that can be applied to the production of other bioanalytical assay critical reagents when platform conjugation approaches fail.

Aim: Pharmacokinetic (PK) characterization and accurate quantification is critical for drug development. The broad potential for antisense oligonucleotide (ASO) drugs to target previously undruggable pathways has resulted in a growing number of ASO therapeutics in development increasing the demand on bioanalysis for PK and safety characterization.

Methods: In this study, a nucleic acid nanorobot (NAN) assay based on toehold mediated strand displacement was for the first time developed for ASO bioanalysis in plasma using a timesaving and simplified 2-step procedure without need for nucleotide extraction. The assay was qualified in two different laboratories to determine precision, accuracy, dilutional linearity, and matrix interferences.

Results: With the ASO driving a programmable cascade of RNA-DNA interaction resulting in release of a fluorophore, the NAN assay generates fluorescent signal proportional to analyte concentration. The assay showed high accuracy and precision across the assay range and high sensitivity. The assay was successfully transferred to a 2^nd laboratory, utilizing different equipment and software, providing evidence of assay robustness and accuracy in mouse plasma.

Conclusions: A NAN assay for ASO PK bioanalysis was for the first time assessed for performance and agreement between laboratories, demonstrating this as a viable additional alternative to current bioanalytical methodologies.

Background: Metabolites were re-mined using MS-DIAL from an SPME dataset preprocessed using AMDIS/Metab R in which data was acquired by GC-MS from schistosomiasis infected (positive) and non-infected (negative)/control samples from school-going children aged between 9 and 12 years in the Okavango Delta region of Botswana. The remining was meant to exhaust scouring for metabolites that could have been overshadowed by Metab R.

Methods: In this study, MS-DIAL open-source freeware was used to preprocess the same dataset to increase the number of identified metabolites and to distinguish positive and control samples.

Results: Ninety-nine (99) metabolites were mined using MS-DIAL in contrast to thirteen identified putatively using Metab R. Both naphthalene and benzophenone, used as surrogate, and internal standards, had greater than 90% match factors using m/z ratios 128 and 105, respectively, for deconvolution with retention time similarity scores of 1000 in all the samples they were added to.

Conclusion: More metabolites were mined using MS-DIAL compared to Metab R thus confirming the preprocessing power of the earlier compared to the latter.