Bioanalysis最新文献

Within the bioanalytical community, the use of blank matrix from preclinical animals for bioanalytical method validation and sample analysis is common practice and required in the context of guidelines for bioanalytical method validation. At the same time, its use has been challenged by the scientific community for decades, since there is ample scientific evidence to allow the use surrogate matrices for this purpose. Nevertheless, legacy and current regulatory thinking continues to be reluctant to allow the use of surrogate matrices in bioanalytical testing except for so-called rare matrices. As part of ongoing discussions in relation to the ICH M10 Guideline, the European Bioanalysis Forum re-challenges the unnecessary use of blank matrices from preclinical animals and believes that, as part of community responsibility and ethical standards and when supported by data, the use of surrogate matrices should become widely accepted. It is in this context that targeted experiments were conducted within the European Bioanalysis Forum to gather additional data and re-open the discussions with all involved and that it should become acceptable to use surrogate matrices wherever possible.

Iron oxide nanozymes (IONzymes) have become fundamental components in various analyte detection methodologies such as colorimetric, electrochemistry, fluorescence and luminescence. Their tunability, stability and the possibility of modification, alongside their ability to mimic the catalytic properties of natural enzymes like peroxidase, render them invaluable in analytical chemistry. This review explores the diverse applications of IONzymes across analytical chemistry, with a particular highlighting on their roles in different detection techniques and their potential in biomedical and diagnostic applications. This information would be valuable for researchers and practitioners in the fields of analytical chemistry, biochemistry, biotechnology and materials science who are interested in applying IONzymes in their work. In essence, this review article on iron oxide nanozymes in analytical chemistry would serve as a valuable resource for researchers, educators and industry professionals, offering insights, guidance and inspiration for further study and application of this promising class of nanomaterials.

Using a straightforward, sensitive and precise liquid chromatographic approach, it is now possible to concurrently measure the amounts of ibuprofen (IBU) and paracetamol (PAR) in human plasma. A µ BondapakTM C18 column (300 mm × 3.9 mm, 15-20 μm) demonstrated acceptable separation when utilizing a mobile phase of 10 mM disodium hydrogen orthophosphate solution and acetonitrile at an 80:20, v/v ratio. The elution was isocratic at room temperature and a flow rate of 1.0 milliliters per minute. The UV detector was set to monitor PAR and IS (tinidazole) for 6.5 min at 254 nm, then IBU for the next 3 min at 220 nm. PAR and IBU showed linearity across the 0.05 to 100 µg/ml concentration range. The precision of the measurements ranged from 98.5% to 105% for PAR and from 95.1% to 102.8% for IBU. The average drug recovery rate was 100% for PAR and 98.9% for IBU. This method was effectively utilized to assess samples from an actual population administered PAR and IBU (325/200 mg) for pharmacokinetic research. The technique employs green and white tools to evaluate their environmental sustainability and efficacy. The suggested strategy was implemented utilizing the Six Sigma method.

Aim: JBD0131, a novel anti-multidrug-resistant tuberculosis (MDR-TB) drug, can target and inhibit the synthesis of mycolic acids, which are crucial components of the cell wall of the Mycobacterium tuberculosis complex. To support the results of this clinical trial in healthy subjects, development of a specific and accurate quantification method for detecting JBD0131 and its metabolite DM131 in human plasma is needed.Materials & methods: Samples with prior added stabilizer were pretreated by protein precipitation method and the extracts were subjected to ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The m/z transitions for the precursor/product ion pairs were 402.1/273 for JBD0131, 333.1/273 for DM131 and 386.1/257 for the internal standard (IS).Results: This method showed good linearity from 1 to 2000 ng/ml for JBD0131 and 0.25 to 500 ng/ml for DM131 and was validated in terms of selectivity, linearity, accuracy, precision, matrix effect, recovery of pretreament and stability.Conclusion: This method was sensitive and specific for measuring the plasma concentrations of JBD0131 and its metabolites. And it was applied for the investigation of the pharmacokinetics of JBD0131 and DM131 in a clinical trial.

The process of developing new drugs in the pharmaceutical industry is both time-consuming and costly, making efficiency crucial. Recent advances in hardware and computational methods have led to the widespread application of computational science approaches in drug discovery. These approaches, including artificial intelligence and molecular simulations, span from target identification to pharmacokinetics research, aiming to reduce the likelihood of failure and present lower costs. Machine learning-based methods predict new applications for developing new drugs based on accumulated knowledge, while molecular simulations estimate interactions between drugs and target proteins at the atomic level based on physical laws. Each approach has its advantages and disadvantages, and they complement each other. As a result, the future of computational science approaches in drug discovery is expected to focus on developing new methodologies that integrate these two techniques to enhance the efficiency of drug discovery.

Assessing the pharmacokinetics of biotherapeutics is essential across all phases of drug development to understand drug exposure. At early stages, platform drug quantification immunoassays offer a versatile method for evaluating exposure for diverse biotherapeutics when specific reagents are not yet available, providing fast data for molecules with common structural features. To ensure clearly defined bioanalytical data, it is essential to conduct interference testing for anti-drug antibodies (ADA) or soluble target (starget), although these assays measure total exposure. In this study, a novel platform immunoassay has been developed that detects a variety of multi-domain drugs with the common structural feature of glycine-serine (G/S) linkers. The assay was successfully qualified and is suitable for early total drug exposure analysis of compounds with G/S linkers. Qualification parameters, including accuracy and precision, were successfully determined, and interference from ADAs and starget was assessed. ADA and starget interference tests showed no impact for one compound, but may affect total drug detection for another. Therefore, it remains recommended to assess the impact of ADAs and starget on assay performance. Overall, the platform G/S linker assay provides a rapid alternative for total exposure analysis in early development when specific assays are unavailable.

Aim: By modifying the Pt electrode with polyvinylpyrrolidone, the distinct oxidation potentials of L and D-serine were markedly increased for both isomers.Methods & results: CV was used for Pt electrode modification with PVP. Twenty cycles was reasonable for Pt modification. K₃[Fe(CN)₆] CV experiments investigated reversibility of the Fe(III) redox reaction and the enhancement of electron transfer at the PVP-Pt electrode. L and D-serine showing an oxidation peak potential at 0.394 V and 0.468 V, respectively. A linear relationship between the anodic oxidation current extracted from the CVs and the standard concentrations of L-serine and D-serine solutions was obtained with R² = 0.99. The dynamic range for D-serine was from 0.5 to 20 mM with a LOD of 0.16 mM, while for L-serine, was from 2.5 to 20 mM with a detection limit of 0.27 mM. Using DPV, the dynamic range was 0.05-1.0 µM for D-serine with a detection limit of 0.0103 µM. The standard deviation ranged from 0.212 to 0.38 across ten determinations per concentration.Conclusion: A separation by 74 mV between the oxidation peaks of L and D-serine was achieved with remarkable enhancement in oxidation current for both isomers. PVP-Pt electrode can detect D-serine in the presence of DL-serine.

In this manuscript, we summarize the discussions and key messages from developed in the e-environment team of the European Bioanalysis forum, which were the basis of a subsequent workshop on cloud applications in a regulated bioanalysis lab environment, hosted by the European Bioanalysis Forum e-environment team at their 16th Open Symposium in Barcelona, Spain in November 2023. The purpose of our discussions is to provide further insight and understanding on the status of having cloud applications implemented in a regulated bioanalysis laboratory and the challenges experienced. The discussions highlight the importance of cross functional collaboration during the entire process of cloud implementation and some of the uncertainties in the different functions' roles and responsibilities.

Over the past years, gene therapeutics have held great promise for treating many inherited and acquired diseases. The increasing number of approved gene therapeutics and developing clinical pipelines demonstrate the potential to treat diseases by modifying their genetic blueprints in vivo. Compared with conventional treatments targeting proteins rather than underlying causes, gene therapeutics can achieve enduring or curative effects via gene activation, inhibition, and editing. However, the delivery of DNA/RNA to the target cell to alter the gene expression is a complex process that involves, crossing numerous barriers in both the extracellular and intracellular environment. Generally, the delivery strategies can be divided into viral-based and non-viral-based vectors. This review summarizes various bioanalysis strategies that support the non-virus-based gene therapeutics research, including pharmacokinetics (PK)/toxicokinetics (TK), biodistribution, immunogenicity evaluations for the gene cargo, vector, and possible expressed protein, and highlights the challenges and future perspectives of bioanalysis strategies in non-virus-based gene therapeutics. This review may provide new insights and directions for the development of emerging bioanalytical methods, offering technical support and a research foundation for innovative gene therapy treatments.

Aim: This study aims to compare the anti-drug antibody (ADA) interference in four pharmacokinetic (PK) assays across different platforms (AlphaLISA, Gyrolab, LC-MS/MS) and to devise a strategy for ADA interference mitigation to improve the accuracy of measured drug in total PK assays.Materials & methods: Spiked test samples, created to achieve different ADA concentrations in human serum also containing an insulin analogue, were analyzed alongside pooled clinical samples using four assays.Results & conclusion: Interference was observed in all platforms. A novel approach using the Gyrolab mixing CD, including acid dissociation in the PK assay, significantly reduced interference and thereby improved relative error from >99% to ≤20% yielding measurements well within the acceptance criteria. Clinical sample results reinforced findings from the test samples.