Yu N, Low J, Macchi F
… +3 more, Vandlen R, Zanghi J, Andrews BT
Bioanalysis
· 2026 Feb · PMID 41983459
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AIM: An interaction observed in an anti-drug antibody (ADA) bridging ELISA between the drug and the anti-digoxin (aDIG) conjugated horseradish peroxidase (HRP) led to increased assay background across several commercial...AIM: An interaction observed in an anti-drug antibody (ADA) bridging ELISA between the drug and the anti-digoxin (aDIG) conjugated horseradish peroxidase (HRP) led to increased assay background across several commercial aDIG-HRP sources. Understanding the source of this interaction will help inform reagent qualities to be considered in production, evaluation, and selection for bioanalytical assays. METHODS AND MATERIALS: This work evaluates commercial sources of aDIG-HRP through bridging ELISA, binding ELISA, SPR, and chromatography to determine what characteristics of aDIG-HRP lead to the interaction observed in a bridging ELISA. RESULTS: Three of four commercial aDIG-HRP sources evaluated exhibit drug binding and size heterogeneity with large (>250 kDa) species. SE-UHPLC of one source localizes this binding capability to the >6 MDa species. Under typical HRP conjugation conditions, HRP can conjugate to itself, and these larger species can bind to drug. CONCLUSION: The conjugation of HRP to aDIG may lead to large species linked to HRP self-conjugation that can interfere with bioassays if not carefully controlled. Therefore, it is important to evaluate commercial reagents for consistency, availability, and critical quality when incorporating a new lot/vendor for better understanding of unexpected assay performance and effectively support reagent and assay life cycle management.
Wang X, Miao H, Liu Z
… +7 more, Sun J, Jiang Z, Ye L, Liang Z, Gao S, Wang Z, Chen W
Bioanalysis
· 2026 Feb · PMID 41958397
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BACKGROUND: SB225002 was a selective non-peptide antagonist of CXC Chemokine Receptor 2 (CXCR2), but its detection method in mass spectrometry and pharmacokinetic (PK) parameters in mouse were not reported. METHODS AND M...BACKGROUND: SB225002 was a selective non-peptide antagonist of CXC Chemokine Receptor 2 (CXCR2), but its detection method in mass spectrometry and pharmacokinetic (PK) parameters in mouse were not reported. METHODS AND MATERIALS: Following methanol-induced protein precipitation and centrifugation, the analyte was separated using an Agilent ZORBAX SB-C18 column (2.1 mm × 100 mm, 3.5 μm, Agilent, MA, USA) with an isocratic mobile phase comprising 25% methanol (phase A) and 75% water containing 0.3% formic acid (phase B). The mobile phase was delivered at a flow rate of 0.3 mL/min. The PK of SB225002 were assessed in a mouse model. RESULTS: SB225002 demonstrated a strong linear relationship in range of 20.00 to 4000.00 ng/mL, with recovery between 96.9% and 99.6%, and matrix effects ranging from 98.9% to 104.6%. SB225002 demonstrated acceptable short-term, long-term, and freeze-thaw stability, with all other items meeting the requirements. The method was utilized to assess PK of SB225002 in mouse, revealing rapid absorption (t: 0.4 ± 0.1 h, mean±SD) and metabolism (t: 3.4 2.6 h), along with significant inter-individual variability. CONCLUSIONS: This study successfully developed and validated a UHPLC-MS/MS method to determine SB225002 in mouse plasma and first reported its pharmacokinetic parameters in mouse, potentially facilitating further clinical research.
Li W, Chen J, Taneja I
… +3 more, Edwards W, Jian W, Geist B
Bioanalysis
· 2026 Feb · PMID 41925326
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BACKGROUND: The blood-brain barrier (BBB) poses challenges for delivering large-molecule therapeutics to the brain. While engineered cross-BBB delivering platforms such as antibodies utilizing receptor-mediated transcyto...BACKGROUND: The blood-brain barrier (BBB) poses challenges for delivering large-molecule therapeutics to the brain. While engineered cross-BBB delivering platforms such as antibodies utilizing receptor-mediated transcytosis (RMT) aim to improve drug delivery, accurate quantization of antibody exposure in the brain is critical. This study investigates pre-analytical factors impacting the analysis of BBB-shuttling antibodies in mice and seeks to bring attention to steps that can be optimized to enhance efficiency and throughput. RESEARCH DESIGN AND METHODS: We evaluated factors such as perfusion, capillary depletion, and freeze-thaw in a humanized mouse model treated with antibodies, with or without a BBB shuttle. Hemoglobin and albumin were investigated as markers for blood contamination. RESULTS: Blood contamination was effectively reduced when cardiac puncture was performed, and perfusion further minimized the contamination. Albumin is a more representative blood contamination marker than hemoglobin for antibody bioanalysis in brain tissue. Capillary depletion did not significantly affect PK analysis under-tested conditions, and no considerable differences were found between frozen and fresh samples. CONCLUSIONS: Careful investigation of these pre-analytical procedures should be conducted to understand their impact. This will facilitate the development of an optimized and efficient workflow for discovery screening.
Li X, Ning Y, Xu J
… +4 more, Gao B, Huang L, Liu Y, Idris A
Bioanalysis
· 2026 Feb · PMID 41915543
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BACKGROUND: Leptomeningeal metastasis (LM) and infectious meningitis often present with similar neurological symptoms, making clinical differentiation challenging. We retrospectively analyzed cerebrospinal fluid (CSF) bi...BACKGROUND: Leptomeningeal metastasis (LM) and infectious meningitis often present with similar neurological symptoms, making clinical differentiation challenging. We retrospectively analyzed cerebrospinal fluid (CSF) biochemistry (lactate (LAC), lactate dehydrogenase (LDH), protein, glucose) in patients with confirmed LM and infectious meningitis (bacterial or viral) to delineate characteristic metabolic profiles. RESEARCH DESIGN AND METHODS: In this single-center retrospective cohort, we included 39 patients with cytology-confirmed LM, 38 with bacterial meningitis (BM), and 40 with viral meningitis (VM). We calculated an LDH × LAC product and compared its diagnostic performance to traditional biochemical markers using ROC analysis and DeLong tests for area under the curve differences. RESULTS: The LDH×LAC index distinguished LM from VM and BM, outperforming the LDH/LAC ratio. Notably, 74.4% of LM patients had low CSF cell counts (≤50 × 10/L) and 66.7% had ≤20% tumor cells, yet LDH×LAC remained highly sensitive even in these low-burden cases. CONCLUSIONS: These findings highlight that certain biochemical signatures can distinguish etiologies whereby high CSF lactate favors bacterial infection or aggressive tumor involvement, whereas normal glucose and low lactate favor viral causes. Our results emphasize distinct CSF signatures in neoplastic compared to infectious meningitis, suggesting metabolic profiling can aid diagnosis.
Cowan KJ, Nelson R, Coddens A
… +17 more, Bloem K, Creed L, Higgins H, Novoa JJ, Jones C, Jyamubandi I, Katterle Y, Klinge M, Krogh-Meibom T, Leatherdale B, Britten C, Bandi DR, Rispens T, Sime K, Videbæk N, Golob M, Timmerman P
Bioanalysis
· 2026 Feb · PMID 41852125
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The European Bioanalysis Forum discusses a recommended shift from the traditional 3-Tier immunogenicity testing approach to a more adaptive immunogenicity testing strategy for biotherapeutics, emphasizing clinical releva...The European Bioanalysis Forum discusses a recommended shift from the traditional 3-Tier immunogenicity testing approach to a more adaptive immunogenicity testing strategy for biotherapeutics, emphasizing clinical relevance, risk-based considerations, assay performance, and collaboration with regulatory authorities to improve the assessment of anti-drug antibodies in therapeutic contexts. The recommendations encourage transitioning toward adaptive 1-Tier or 2-Tier combined approaches tailored to the molecule context-of-use. The new strategy prioritizes the clinical impact of anti-drug antibodies over mere incidence, advocating careful consideration of assay characteristics during assay development. The European Bioanalysis Forum continues to call for published case studies, industry cooperation, and proactive dialogue with regulatory authorities to challenge the traditional 3-Tier approach.
Fernández-Metzler C, Quadrini KJ, Amaravadi L
… +9 more, Cape S, Green J, Hays A, Kamphorst JJ, Laxmanan S, Pandey A, Qiu X, Rajapaksha RD, Hassanein M
Bioanalysis
· 2026 Jan · PMID 41804680
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Pharmacodynamic (PD) biomarkers provide crucial insights into a drug's mechanism of action (MoA) and efficacy by measuring its effects on biological targets within an organism. PD biomarkers can be proximal (e.g. recepto...Pharmacodynamic (PD) biomarkers provide crucial insights into a drug's mechanism of action (MoA) and efficacy by measuring its effects on biological targets within an organism. PD biomarkers can be proximal (e.g. receptor occupancy, enzyme inhibition) or distal (e.g. downstream pathway modulation) to the biological target. In drug development, PD biomarkers are essential for monitoring patient response, assessing therapeutic efficacy, optimizing dosage strategies, and streamlining the drug development process by informing go/no-go decisions. In personalized medicine, PD biomarkers enable tailored treatments based on individual responses, enhancing both effectiveness and safety. Sound bioanalytical strategies and rigorous assay validation practices are key for successful integration of PD biomarkers into clinical trials. This paper outlines the bioanalytical and assay considerations for developing and validating informative PD biomarker assays and their use in drug development.
Zhang B, Zhang L, Liu Y
… +5 more, Hu W, Diao R, Xing L, Tao Y, Shen L
Bioanalysis
· 2026 Feb · PMID 41797599
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INTRODUCTION: Antibody-drug conjugates (ADCs) represent a cutting-edge approach in cancer therapy, with monomethyl auristatin E (MMAE) frequently used as a payload in ADC development. We have established a novel bioanaly...INTRODUCTION: Antibody-drug conjugates (ADCs) represent a cutting-edge approach in cancer therapy, with monomethyl auristatin E (MMAE) frequently used as a payload in ADC development. We have established a novel bioanalytical method characterized by high sensitivity, accuracy, and efficiency for quantifying MMAE in cynomolgus monkey plasma. METHODOLOGY: MMAE was extracted using liquid-liquid extraction (LLE) and quantified by Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS). Results: The method exhibited excellent linearity across a concentration range of 5 to 3000 pg/mL ( ≥ 0.99) in cynomolgus monkey plasma, it fulfilled precision (CV ≤ 15%, 20% for lower limit of quantification (LLOQ)) and accuracy (83.98-112.75%) criteria according to meeting validation requirements per regulatory bioanalytical validation guidelines. CONCLUSION: This validated method is instrumental in exploring the in vivo pharmacokinetic profile of ADC and elucidating their exposure-response dynamics.
Bioanalysis
· 2026 Jan · PMID 41777170
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Advances in analytical techniques now enable biomarker detection from sample volumes as small as one microliter, reducing the need for large-volume specimen collection. This capability has supported the development of lo...Advances in analytical techniques now enable biomarker detection from sample volumes as small as one microliter, reducing the need for large-volume specimen collection. This capability has supported the development of low-volume blood collection devices that allow frequent, longitudinal sampling outside traditional healthcare facilities, promoting decentralized and patient-centered sampling (PCS). The development of user-friendly, at-home sampling devices with reduced invasiveness and minimized sampling discomfort further enhances participant recruitment and retention in long-term longitudinal studies. These longitudinal studies are essential for tracking disease progression, uncovering molecular mechanisms underlying disease, and gaining insights into how external stimuli, such as diet, medication and physical activity, affect physiological responses. In this review, we highlight current low-volume sampling technologies and their role in longitudinal PCS environments, alongside biomarker detection assays for small-volume specimens.
Yang N, Liu J, Mu K
… +4 more, Liang A, Yin G, Wang X, Liu R
Bioanalysis
· 2026 Jan · PMID 41773027
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AIM: Sodium carboxymethylcellulose acts as both a pharmaceutical excipient (interacting with drugs to affect metabolism) and an active substance (stimulating intestinal peristalsis via aqueous polymeric colloids). To cla...AIM: Sodium carboxymethylcellulose acts as both a pharmaceutical excipient (interacting with drugs to affect metabolism) and an active substance (stimulating intestinal peristalsis via aqueous polymeric colloids). To clarify its fate and advance sodium carboxymethylcellulose-based adjuvant delivery, a targeted analysis was needed, using dextromethorphan hydrobromide as a model drug. RESEARCH DESIGN AND METHOD: A comprehensive LC-MS approach was established: UHPLC-Q TRAP MS/MS (quantitative) and HPLC-Q TOF MS/MS (qualitative) were used to study sodium carboxymethylcellulose pharmacokinetics, distribution, metabolism, and excretion in male Wistar rats, with/without co-administered dextromethorphan hydrobromide. RESULTS: The distribution of sodium carboxymethylcellulose is expedited to organs characterized by augmented blood flow velocities, including the spleen, liver, and heart. And about 66.4% sodium carboxymethyl cellulose was removed from plasma within 36 h. Results also showed that the excretion level of sodium carboxymethyl cellulose is less than the dosage administered, and almost all glycosidic bonds had been hydrolyzed in its process. CONCLUSION: The findings provide complete description of sodium carboxymethylcellulose fate, establishing a novel framework that facilitates the extensive utilization of this pharmaceutical excipient in the process of drug formulation development.
Bioanalysis
· 2026 Jan · PMID 41762632
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BACKGROUND: Sepsis-induced immunosuppression is a significant contributor to mortality, with the programmed cell death protein 1 and its ligand (PD-1/PD-L1) pathway playing a central role in T-cell exhaustion. While thei...BACKGROUND: Sepsis-induced immunosuppression is a significant contributor to mortality, with the programmed cell death protein 1 and its ligand (PD-1/PD-L1) pathway playing a central role in T-cell exhaustion. While their dynamics as biomarkers for monitoring treatment response in specific infections, such as Enterobacteriaceae-induced sepsis, remain unclear. MATERIALS AND METHODS: This study enrolled 63 patients with Enterobacteriaceae septicemia and 59 healthy controls. Serum levels of sPD-1/sPD-L1 were quantified at admission (Day 0) and on Days 3, 7, and 14. Patients were stratified into responders and non-responders based on clinical outcomes. RESULTS: At Day 0, sepsis patients had significantly higher levels of sPD-1 and sPD-L1 compared to healthy controls ( < 0.001). Longitudinal analysis revealed that patients who responded favorably to treatment exhibited a significant progressive decline in both biomarkers over the 14-day period. In contrast, non-responders maintained persistently elevated levels. Furthermore, initial sPD-1/sPD-L1 concentrations positively correlated with SOFA score, lactate level and PCT level. Furthermore, the analysis showed that the antibiotics β-lactam/β-lactamase inhibitor and carbapenem might not directly influence the sPD-1/sPD-L1 levels. CONCLUSION: Serum sPD-1 and sPD-L1 levels are significantly elevated in Enterobacteriaceae sepsis and dynamically decreased in patients responding to therapy. These findings underscore their potential as valuable prognostic biomarkers for monitoring immune reconstitution and treatment efficacy.
Bioanalysis
· 2026 Jan · PMID 41761838
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INTRODUCTION: Immunogenicity remains the principal constraint on the efficacy, safety, and re-dosing of adeno-associated virus (AAV) and lipid nanoparticle (LNP) - mRNA therapeutics. Clinical decision-making and comparab...INTRODUCTION: Immunogenicity remains the principal constraint on the efficacy, safety, and re-dosing of adeno-associated virus (AAV) and lipid nanoparticle (LNP) - mRNA therapeutics. Clinical decision-making and comparability between programs are hampered by heterogeneous test formats and matrix effects. AREAS COVERED: This narrative review integrates 2023-2025 bench, translational, clinical, and regulatory evidence. Studies on binding, neutralizing, and cellular/innate tests, anti-PEG epidemiology, complement biology, and ultrasensitive digital immunoassays were identified through searches of PubMed, Web of Science, Google Scholar, and regulatory archives. We propose a panel with tiers: Tier-0 pre-screening using isotype-resolved anti-PEG and total anti-capsid; Tier-1 standardized cell-based transduction inhibition with an optional constant-serum-concentration design to reduce dilution-induced matrix bias; Tier-2 mechanistic cellular and innate panels (e.g. complement split products, cytokines, and capsid-specific T-cell assays); and Tier-3 attomolar digital assays for critical biomarkers. FUTURE PERSPECTIVE: By implementing this harmonized, matrix-aware panel, which is presented as a fit-for-purpose analytical best-practice proposal rather than an evidence-validated clinical standard, screening-failure bias can be minimized, eligibility criteria can be stabilized, and connections between assay results and dosing, monitoring, and re-dosing/deferral may be strengthened.
Karlsborn T, Stenbratt CL, Nain-Perez A
… +1 more, Hölttä M
Bioanalysis
· 2026 Jan · PMID 41755553
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BACKGROUND: Approaches such as liquid-liquid extraction and two-step liquid-liquid extraction combined with SPE has been widely used for extraction of oligonucleotide therapeutics. It has been more challenging to develop...BACKGROUND: Approaches such as liquid-liquid extraction and two-step liquid-liquid extraction combined with SPE has been widely used for extraction of oligonucleotide therapeutics. It has been more challenging to develop a generic one-step SPE method that can be used across multiple tissue types. MATERIALS & METHODS: An SPE method was developed for extraction of oligonucleotide therapeutics from tissue homogenates utilizing an ASO internal standard and a siRNA analyte. The method was fully automated using a liquid handler and an automated SPE station. Quantification of the siRNA antisense was performed using LC-MS, with calibration samples prepared in a surrogate matrix. RESULTS: The fully automated SPE method enabled quantification of the siRNA antisense across nine different tissues from three different species. The SPE method utilizing surrogate matrix calibration was compared to liquid-liquid extraction by analyzing in-vivo samples using both workflows, showing good agreement between the two different extraction methods. CONCLUSIONS: This manuscript presents a one-step SPE method greatly reducing blank tissue requirements and sample preparation workload by enabling use of surrogate matrix calibration and full automation. The presented SPE method simplifies and reduces analytical time in bioanalysis of tissue biodistribution studies and adheres to the principles of 3R.
Mottagi Khosrowshahi H, Abolhasani J, Afshar Mogaddam MR
… +2 more, Ghorbani Kalhor E, Ghasemi E
Bioanalysis
· 2026 Jan · PMID 41755504
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BACKGROUND: The therapeutic bioanalytical challenge activity of tyrosine kinase inhibitors (TKIs) can be assessed by measuring them in biological samples such as plasma and serum. Determining these compounds requires an...BACKGROUND: The therapeutic bioanalytical challenge activity of tyrosine kinase inhibitors (TKIs) can be assessed by measuring them in biological samples such as plasma and serum. Determining these compounds requires an efficient sample before the analysis to minimize matrix effects and enhance preconcentration. This study presents the use of a composite of layered double hydroxide (LDH) and metal organic framework (MOF) composite for extracting and preconcentrating of sunitinib, imatinib, and dasatinib. METHODOLOGY: The proposed composite was synthesized through the in-situ growth of MIL-101(Cr)-NO MOF on the MgFe-LDH surface. The extraction process involves adding 5 mg of LDH/MIL-101(Cr)-NO to the plasma solution containing the analytes, under nitrogen blowing. Centrifugation separated the analyte-loaded sorbent, which was then eluted with 125 μL of mobile phase. The organic phase was used subsequent high performance liquid chromatography (HPLC) - photo diode array detector (PDA) analysis. RESULTS: Under optimum conditions, the developed method demonstrates favorable extraction recoveries (66-74%), low limits of detection (LOD) and quantification (LOQ) (0.46-0.69 ng mL and 1.5-2.2 ng mL, respectively), wide linear ranges (LRs) (1.5-200 ng mL) with high coefficients of determination (0.997-0.999) and proper repeatability (RSD ≤ 6.6%). CONCLUSIONS: Notably, the method provides the use of minimal amounts of sorbent and solvents, short extraction time, no need for complicated instruments, and high performance in monitoring anti-cancer drugs in biological samples.
Tsai PC, Condos T, Devlin J
… +7 more, Chaitanya K, Geyer Prinslow E, Dong Y, Chao Y, Wu B, Luo J, Jiang Y
Bioanalysis
· 2026 Jan · PMID 41742824
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AIM/BACKGROUND: Anti-idiotype antibodies (anti-IDs) targeting therapeutic antibodies play a crucial role in the development of pharmacokinetic (PK), antidrug antibody (ADA), and neutralizing antibody (NAb) assays during...AIM/BACKGROUND: Anti-idiotype antibodies (anti-IDs) targeting therapeutic antibodies play a crucial role in the development of pharmacokinetic (PK), antidrug antibody (ADA), and neutralizing antibody (NAb) assays during preclinical/clinical phases. However, the generation of fit-for-purpose anti-IDs poses significant challenges, especially for NAb assay development, which requires high-affinity anti-IDs to be effectively competed with the target for its binding to the drug. Our preliminary results from NAb screenings revealed that all 19 anti-IDs generated from a mouse immunization campaign in conjunction with a hybridoma platform failed to demonstrate the ability to effectively inhibit the binding of therapeutic tri-specific mAb1 to the target arm protein 1. METHOD/RESULTS: We subjected the selected parental clone to affinity maturation utilizing phage display technology aimed at enhancing binding affinity and mitigating matrix interference. Following four rounds of panning, our assessments via enzyme-linked immunosorbent assay (ELISA) and biolayer interferometry (BLI) analysis demonstrated considerable enhancements in the binding affinity of 11 selected clones. Notably, three candidates further exhibited strong neutralizing capabilities to the drug target arm protein 1 within the matrix for NAb assays. CONCLUSION: Affinity-improved anti-IDs enabled the development of a robust and fit-for-purpose competitive ligand-binding NAb assay.
Kameyama T, Imai S, Mitamura R
… +3 more, Niwa M, Yamada T, Kusano K
Bioanalysis
· 2026 Jan · PMID 41711019
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BACKGROUND: LC-MS/MS is widely used for the quantification of phosphorodiamidate morpholino oligomers, but its low sensitivity is a problem. In addition, solid-phase extraction, a pretreatment method generally used for L...BACKGROUND: LC-MS/MS is widely used for the quantification of phosphorodiamidate morpholino oligomers, but its low sensitivity is a problem. In addition, solid-phase extraction, a pretreatment method generally used for LC-MS/MS analysis, results in low recovery when applied to phosphorodiamidate morpholino oligomers. Therefore, a highly sensitive bioanalytical method for the determination of phosphorodiamidate morpholino oligomers using LC-MS/MS with improved pretreatment is desired. RESULTS: Protein precipitation was applied to LC-MS/MS analysis for viltolarsen, as a typical phosphorodiamidate morpholino oligomer, in plasma, and PPT with MeOH achieved high recoveries (>90%). Reversed-phase liquid chromatography (RPLC) with gradient elution improved ion suppression compared to hydrophilic interaction liquid chromatography. A combination of PPT and RPLC achieved higher sensitivity (lower limit of quantification, 1 ng/mL) than those of approved phosphorodiamidate morpholino oligomers (10 to 20 ng/mL in their clinical reviews), and the between-run accuracy and precision ranged from 97.1% to 109% and 2.56% to 10.5%, respectively. This method was also shown to be applicable to another phosphorodiamidate morpholino oligomer, ψM23D (+07-18). CONCLUSION: A simple and highly sensitive bioanalytical method for the determination of viltolarsen was established by optimizing RPLC and PPT, and the method was successfully applied to a phosphorodiamidate morpholino oligomers with a different sequence.
Xu W, Luo L, Matta M
… +2 more, Helmy R, Vazvaei-Smith F
Bioanalysis
· 2026 Jan · PMID 41708146
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Antibody-drug conjugates (ADCs) have reemerged as a transformative therapeutic modality, with over a dozen approvals and hundreds of candidates in clinical development. Historically, ADC pharmacokinetic (PK) evaluation r...Antibody-drug conjugates (ADCs) have reemerged as a transformative therapeutic modality, with over a dozen approvals and hundreds of candidates in clinical development. Historically, ADC pharmacokinetic (PK) evaluation required measuring all constituent components - total antibody (TAb), conjugated antibody (ADC), or conjugated payload, free payload, and major metabolites. This comprehensive approach led to a significantly higher bioanalytical (BA) investment, increased patient blood volume requirements, and far more resources than those needed for monoclonal antibody (mAb) therapeutics or small molecule drugs. In 2024, the US Food and Drug Administration (FDA) issued guidance encouraging a more streamlined, data-driven approach to ADC PK assessment in clinical development, including the potential to omit certain analytes when scientifically justified. While this guidance represents a pivotal shift, adoption across the industry has been slow, and few examples of successful assay reduction have been publicly shared. In this perspective, we examine the scientific and regulatory rationale for simplifying ADC PK strategies. We also propose a pragmatic framework for reducing assays - grounded in clinical relevance, early-phase data, and platform knowledge - that seeks to reduce complexity without compromising patient safety or data integrity.
Emrich T, Schick E, Wolfert A
… +10 more, Meir J, Guo X, Kubalec P, Stubenrauch KG, Ockenga K, Reis-Dybowski M, Vauleon S, Heinrich J, Barfield M, Staack RF
Bioanalysis
· 2026 Jan · PMID 41705351
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AIMS: This study assessed the automated cobas e411 immunoanalyzer as a high-efficiency alternative to manual ELISA for pharmacokinetic and pharmacodynamic analyses. Key objectives included reducing manual resource burden...AIMS: This study assessed the automated cobas e411 immunoanalyzer as a high-efficiency alternative to manual ELISA for pharmacokinetic and pharmacodynamic analyses. Key objectives included reducing manual resource burdens and minimizing analyst-dependent variability while achieving analytical performance comparable or superior to established ELISA standards. MATERIALS & METHODS: To achieve GLP and FDA 21 CFR Part 11 and 58 compliance, a dedicated "Elecsys system", comprising the off-the-shelf cobas e411 platform and an in-house developed software package (CoLiBri) for instrument control and LIMS connection was subjected to Computerized System Validation (CSV). The Elecsys system's analytical performance was compared directly to ELISA. Robustness was further assessed through clinical sample analysis and free analyte quantification in biological matrices. RESULTS: The cobas e411 demonstrated comparable sensitivity and precision to ELISA while offering a significantly wider dynamic range due to electrochemiluminescence readout. Automation minimized human error and enabled high-throughput singlicate analysis, a capability unattainable with manual methods. Validation of numerous non-clinical and clinical assays and routine sample analyses confirmed platform's efficiency and ability to provide rapid turnaround for bioanalysis. CONCLUSIONS: The Elecsys system is a robust, GLP-compliant platform that streamlines bioanalytical workflows. By automating labor-intensive steps, it significantly enhances data quality and efficiency compared to traditional manual ligand-binding assays.
Wojcik J, Qian M, Rosenbaum AI
… +63 more, Maes E, Xue Y, Kochansky C, Boer J, Chen L, Descloux S, Garofolo F, Johnson J, Kaur S, Lassman M, McGregor J, O'Callaghan L, Sikorski T, Wan K, Wang S, Zheng N, Zhang G, Hoover M, Wang J, Yuan L, Ji Q, Jones B, Boyanapalli R, Chen J, Dobson C, Dong L, Ferrari L, Fu Y, Gong C, Ivanova D, Li J, Licea-Perez H, Liu A, Tang X, Tao L, Tweed J, Eddy J, Francis J, Kavetska O, Araya M, Foley T, Andisik M, Belfast M, Borbridge L, Cho SJ, Colligan L, Dasgupta A, Edmison A, Fandozzi C, Galliccia F, Kim YJ, Lu Y, Massip AM, Shakleya D, Tampal N, Neto JT, Thomas J, Wang J, Wang L, Wang Y, Whale E, Wiberg M, Yang L
Bioanalysis
· 2025 Dec · PMID 41685725
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The 19 Workshop on Recent Issues in Bioanalysis (19 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 conv...The 19 Workshop on Recent Issues in Bioanalysis (19 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 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 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 1) covers in the Part 1A the recommendations on Mass Spectrometry Assays and Regulated Bioanalysis/BMV and in Part 1B the Regulatory Inputs on these topics. Part 2 (Biomarkers/BAV, IVD/CDx, Ligand-Binding Assays and Cell-Based Assays) and Part 3 (Gene Therapy, Cell therapy, Vaccines and Biotherapeutics Immunogenicity) are published in volume 18 of Bioanalysis, issues 1 and 2 (2026), respectively.
Bioanalysis
· 2026 Jan · PMID 41684060
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BACKGROUND: PrC-210 is a direct-acting reactive oxygen species (ROS) scavenger. It is active when administered orally, IV, or subcutaneously. Its profound efficacy in preclinical models makes it a candidate to prevent hu...BACKGROUND: PrC-210 is a direct-acting reactive oxygen species (ROS) scavenger. It is active when administered orally, IV, or subcutaneously. Its profound efficacy in preclinical models makes it a candidate to prevent human ROS-induced i) ionizing radiation toxicities, ii) neurodegenerative disease, and iii) organ ischemia-reperfusion injuries. RESEARCH DESIGN AND METHODS: A simple, highly sensitive, specific LC-MS/MS method was developed to measure the PrC-210 molecule in mouse plasma and tissue homogenates. To prevent oxidative degradation or disulfide formation of the PrC-210 thiol form, its free sulfhydryl group was protected by derivatization with N-ethylmaleimide (NEM), which produced the stable, quantifiable, PrC-210-NEM thioether conjugate. The PrC-210-NEM conjugate, and acyclovir used as an internal standard, were extracted from plasma or tissue homogenate with acetonitrile/0.1% formic acid. RESULTS: The reconstituted dried extracts were chromatographed and then monitored using a triple quadrupole spectrometer operating in the positive ion spray ionization mode. The method was validated over the concentration range of 7.5 nM-5000 nM. Inter- and intra-assay precision and accuracy of PrC-210-NEM quantitation were better than 10%. The limit of PrC-210-NEM quantitation was 2.5 nM. CONCLUSIONS: The method was applied to measure plasma and tissue homogenate concentrations of PrC-210 in over 126 mice administered either oral, itraperitoneal, or subcutaneous PrC-210 doses.