A multilaboratory validation study of LC/MS biomarker assays for three lysophosphatidylcholines

Bioanalysis ◽  
2021 ◽  
Author(s):  
Rika Ishikawa ◽  
Kosuke Saito ◽  
Tsuyoshi Matsumura ◽  
Koji Arai ◽  
Saki Yamauchi ◽  
...  
Keyword(s):  
Clinical Application ◽  
Validation Study ◽  
Calibration Curve ◽  
Rat Plasma ◽  
Relative Accuracy ◽  
Acceptance Criteria ◽  
Assay Validation ◽  
Regulatory Documents ◽  
Validation Parameters ◽  
Fit For Purpose

Aim: Although the fit-for-purpose approach has been proposed for validation procedures and acceptance criteria for biomarker assays, practical biomarker assays to facilitate clinical application and regulatory documents on biomarker assays remain limited. Materials & methods: We assigned six independent laboratories and selected three lysophosphatidylcholines (LPCs): LPC(16:0), LPC(18:0) and LPC(18:1) as model biomarkers. Using LC–MS, the following key validation parameters were evaluated: calibration curve, carryover, parallelism, precision and relative accuracy and these values were similar among all laboratories. Further, we determined LPC levels in six lots of rat plasma at unknown concentrations and compared them among the laboratories. Conclusion: Our multilaboratory validation and reproducibility data are useful for the development of future biomarker assay validation procedures, as well as regulatory documents.

P0301FIT FOR PURPOSE VALIDATION OF A CLINICAL ASSAY FOR THE DETECTION OF SERUM LEVELS OF GALACTOSE-DEFICIENT IMMUNOGLOBULIN A1 (GD-IGA1) IN PATIENTS WITH IGA NEPHROPATHY (IGAN)

2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Laura Sponton ◽  
Hulin Jin ◽  
Markus Fluck ◽  
Yusuke Suzuki ◽  
Amy Kao
Keyword(s):  
Quantitative Determination ◽  
Human Serum ◽  
Iga Nephropathy ◽  
Calibration Curve ◽  
Clinical Studies ◽  
Serum Samples ◽  
Freeze Thaw ◽  
Healthy Donors ◽  
Validation Parameters

Abstract Background and Aims Analysis of serum or plasma from patients with IgA nephropathy (IgAN) has confirmed the presence of elevated levels of circulating immune complexes containing Gd-IgA1 (Czerkinsky 1986). New sensitive and reasonably specific noninvasive tests are emerging to guide the therapeutic strategy that is applicable to all stages of IgAN (Suzuki 2014). Here we are reporting the fit for purpose validation of an ELISA method for the quantitative determination of Gd-IgA1 in human serum samples to support biomarker investigations in clinical studies of Merck KGaA, Darmstadt. Method The assay was developed based on a commercially available immunoassay kit. The dynamic range of the calibration curve was determined from 1.56 ng/mL (LLOQ) to 100 ng/mL (ULOQ). With a minimum required dilution of 200-fold and standard assay volume of 50.0 μL, the range of the method in matrix was from 312 ng/mL to 20, 000 ng/mL. In assay validation phase, multiple validation parameters were evaluated, which included minimum required dilution (MRD), calibration curve, matrix effect, Intra- & Inter run accuracy & precision, selectivity, and parallelism. Additional validation parameters include sample stability (short/long term, freeze-thaw) and batch-to-batch comparison. Results All samples measured for intra & Inter - assay precision, accuracy, fulfilled the specifications according to the acceptance criteria. The selectivity was assessed using blank serum matrix from 10 individuals: the result indicated that matrix components in serum did not interfere with the detection of Gd-IgA1. Parallelism assessment was performed successfully for both samples from healthy donors and IgAN patient samples up to dilution factor (DF) 3200 (serum samples from healthy donors were determined up to DF 1600). All DF-corrected results within the assay range were determined with %CV ≤ 30.0%. Batch to batch comparison was assessed successfully based on the known shelf life of the kit. Short term stability using QC samples were given for up to 24hrs at room temperature. Freeze-thaw stability was given for up to 3 cycles at -20°C±5°C and -75°C±15°C. The investigations were performed according to general guidelines for method validation and applicable regulations. The results of investigated validation parameters fulfilled the requirements and recommendations, generally accepted for bioanalytical projects. Conclusion The present validation qualified the method for the quantitative determination of Gd-IgA1 in human serum samples from clinical studies.

scholarly journals Cross-Platform Analysis of HIV-1 RNA Data Generated by a Multicenter Assay Validation Study with Wide Geographic Representation

2012 ◽  
Vol 50 (8) ◽  
pp. 2737-2747 ◽  
Author(s):  
C. Jennings ◽  
B. Harty ◽  
S. Granger ◽  
C. Wager ◽  
J. A. Crump ◽  
...  
Keyword(s):  
Validation Study ◽  
Assay Validation ◽  
Cross Platform ◽  
Hiv 1

An Inter-Laboratory Idiotype Assay Validation Study

Lupus ◽  
1992 ◽  
Vol 1 (5_suppl) ◽  
pp. 317-322 ◽  
Author(s):  
Ewa Cairns ◽  
Susan Andrejchyshyn ◽  
Joyce Rauch
Keyword(s):  
Validation Study ◽  
Assay Validation

scholarly journals Adalimumab and anti-adalimumab LISA-TRACKER immunoassays performance criteria for therapeutic drug monitoring of adalimumab-amgen biosimilar (ABP501)

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Fabien Francois ◽  
Loubna Naimi ◽  
Xavier Roblin ◽  
Anne-Emmanuelle Berger ◽  
Stephane Paul
Keyword(s):  
Human Serum ◽  
Dynamic Range ◽  
Polyclonal Antibodies ◽  
Performance Criteria ◽  
Clinical Samples ◽  
Therapeutic Drug ◽  
Serum Samples ◽  
Acceptance Criteria ◽  
Sample Stability ◽  
Validation Parameters

Abstract Background ABP501 is a biosimilar to Reference Adalimumab (HUMIRA®) produced by AMGEN. Adalimumab (ADA) has a marketing authorization for Crohn's disease, ulcerative colitis and other inflammatory or autoimmune diseases. The aim of this study was to evaluate the LISA-TRACKER assays developed by Theradiag (France), for the monitoring of ABP501 and anti-ABP501 antibodies in human serum. Results 68 ABP501 clinical samples were measured with the LISA TRACKER Duo Adalimumab assay. LISA TRACKER has been validated as suitable for quantification of ABP501 in human serum samples. Accuracy of the LISA-TRACKER was measured using 3 human serum matrices spiked with known levels of biosimilar, 3 levels spanning the dynamic range. Percentages of recovery were ranged from 90 to 120% for biosimilar batch1, and between 93 and 105% for biosimilar batch2. The acceptance criteria (CV < 20%) were met for intra-run (from 3.8 to 16.5%) and inter-run imprecision (from 4.4 to 13.9%) including the two batches. All results were comprised within ± 20% from results, obtained with the kit and sample unexposed in order to evaluate stability of the sample, stability of the kit and consistency of the results. In any case, but two, all percentages of inhibition were > 50% for specificity. Specificity was tested with Biosimilar spiked samples, Biosimilar with Humira® spiked samples, and clinical samples from patients treated with adalimumab biosimilar. All of these samples were spiked with polyclonal antibodies directed against Humira®. Specificity inhibition and specificity detection steps were also part of the validation parameters. Reagents made with ABP501 gave similar results than reagents made with Humira® meeting acceptance criteria. Conclusions LISA-TRACKER ADA and anti-ADA assays are reliable for the monitoring of patients treated with ABP501.

scholarly journals NOVEL UV SPECTROPHOTOMETRIC METHOD FOR THE DETERMINATION OF TERIFLUNOMIDE IN TABLET DOSAGE FORM

2019 ◽  
pp. 81-84
Author(s):  
VISHWAS T. S. ◽  
GURUPADAYYA B. M.
Keyword(s):  
Linear Relationship ◽  
Spectrophotometric Method ◽  
Calibration Curve ◽  
Dosage Form ◽  
Current Work ◽  
Validation Parameters ◽  
Line Equation ◽  
Tablet Dosage ◽  
Ich Guideline

Objective: The current work is intended towards the development of a novel, simple, and precise UV spectrophotometric method for the estimation of teriflunomide (TEF) present in the marketed formulation. Methods: Acetonitrile was used as asolvent and the absorbance of the drug was measured at the absorbance maxima of TEF, UV 284 nm. Results: Calibration curve plotted in concentration range 5-10 µg /ml exhibited excellent linear relationship with line equation y = 0.0858x-0.0223 and r2 value of 0.9996. The method was found to comply all the validation parameters as per the ICH guideline indicating the sensitivity of the method towards analyte. Conclusion: The method can be used satisfactorily for the routine analysis of TEF present in marketed formulation.

scholarly journals Enhanced antipneumococcal antibody electrochemiluminescence assay: validation and bridging to the WHO reference ELISA

Bioanalysis ◽  
2020 ◽  
Vol 12 (19) ◽  
pp. 1363-1375
Author(s):  
Katrina M Nolan ◽  
Yuhua Zhang ◽  
Joseph M Antonello ◽  
Adrienne H Howlett ◽  
Cyrille J Bonhomme ◽  
...  
Keyword(s):  
Clinical Trial ◽  
Clinical Trials ◽  
Conjugate Vaccine ◽  
Threshold Value ◽  
Phase Iii ◽  
Capsular Polysaccharides ◽  
Acceptance Criteria ◽  
Phase Iii Clinical Trial ◽  
Assay Validation ◽  
Clinical Trial Program

Aim: To re-optimize the pneumococcal (Pn) electrochemiluminescence (ECL) assay and to validate and bridge the enhanced assay to the WHO ELISA, to support the Phase III clinical trial program for V114, a 15-valent Pn conjugate vaccine. Materials & methods: The Pn ECL assay was re-optimized, validated and formally bridged to the WHO ELISA. Results: The enhanced Pn ECL assay met all prespecified validation acceptance criteria and demonstrated concordance with the WHO ELISA. The corresponding threshold value remains at 0.35 μg/ml for all 15 serotypes. Conclusion: The enhanced Pn ECL assay has been validated for the measurement of antibodies to 15 Pn capsular polysaccharides and is concordant with the WHO ELISA, supporting its use in clinical trials.

scholarly journals Simultaneous Determination of Seven Active Components in Rat Plasma by UHPLC-MS/MS and Application to a Quantitative Study after Oral Administration of Huang-Lian Jie-Du Decoction in High Fat-Induced Atherosclerosis Rats

2019 ◽  
Vol 2019 ◽  
pp. 1-12 ◽  
Author(s):  
Li Jiang ◽  
Yanling Xiong ◽  
Lanbin Yu ◽  
Yu Chen ◽  
Qiyun Zhang ◽  
...  
Keyword(s):  
Oral Administration ◽  
Simultaneous Determination ◽  
Quantitative Study ◽  
High Performance ◽  
Rat Plasma ◽  
Dependent Manner ◽  
High Fat ◽  
Active Components ◽  
Validation Parameters

Huang-Lian Jie-Du decoction (HLJDD) has been used to treat cardiovascular and cerebrovascular disease for many years in China. Currently, the determination of effect components in HLJDD is focusing either on the formula or on the extract, while quantification of that in biological samples is scarce, especially simultaneous determination of multicomponent. In this paper, a rapid, specific, and sensitive ultra-high performance liquid chromatography-tandem mass spectrometry method was developed and fully validated for the simultaneous determination of seven main active constituents, i.e., baicalin, baicalein, wogonoside, wogonin, berberine, palmatine, jatrorrhizine in rat plasma. The method was also successfully applied to a quantitative study after oral administration of HLJDD at different doses of 1.5, 3, and 6 g/kg body weight to high fat-induced atherosclerosis rats. The analytes were detected by ESI source and multiple reactions monitoring (MRM) using positive scanning mode. The blood was collected from the abdominal aorta of rats at predetermined time and preprepared with icariin and tetrahydropalmatine as internal standards (IS). Sample preparation was achieved by protein precipitation (PPT). The validation parameters (linearity, sensitivity, intra-/interday precision and accuracy, extraction recovery, and matrix effect) were within acceptable ranges, and biological extracts were stable during the entire storing and preparing process. And the result of determination of HLJDD-containing plasma, baicalin, baicalein, wogonoside, and wogonin could be highly detected in a dose-dependent manner while berberine, jatrorrhizine, and palmatine were determined in a very low level and in a dose-independent mode. Thus, the established method was sensitive enough and successfully applied to the determination of seven effective components in plasma taken from 24 high fat-induced atherosclerosis rats after oral administration of three dosages of HLJDD.

scholarly journals Pest occurrence model in current climate – validation study for European domain

2013 ◽  
Vol 61 (1) ◽  
pp. 205-214 ◽  
Author(s):  
Eva Svobodová ◽  
Miroslav Trnka ◽  
Martin Dubrovský ◽  
Daniela Semerádová ◽  
Josef Eitzinger ◽  
...  
Keyword(s):  
Validation Study ◽  
Cydia Pomonella ◽  
Rhopalosiphum Padi ◽  
Distribution Patterns ◽  
Lobesia Botrana ◽  
Sensitivity Analyses ◽  
Oulema Melanopus ◽  
Model Parameterization ◽  
Current Climate ◽  
Validation Parameters

The present study yields detail validation of the pest occurrence models under current climate in wide European domain. Study organisms involve Cydia pomonella, Lobesia botrana, Ostrinia nubilalis, Leptinotarsa decemlineata, Oulema melanopus, Rhopalosiphum padi, and Sitobion avenae. Method used in this study belongs to the category climate matching (CLIMEX model) allowing the estimation of areas climatically favourable for species persistence based on the climatic parameters characterising the species development. In the process of model validation parameters were iteratively tested and altered to truly describe the pest presence. The modelled pests presence was verified by comparison of the observed pests occurrence with the number of generations in given modelled area. The notable component of the model parameterization was the sensitivity analyses testing the reaction of species development on changing meteorological items. Parameterization of the factors causing distribution patterns of study species was successful and modelled potential distributions of species correspond well to known core distribution areas for all of these species. This validation study is intended as an initial for forthcoming studies focused on the estimation of geographical shifts of selected pests in the conditions of climate change within the Europe.

scholarly journals Adalimumab and Anti-adalimumab Lisa-tracker Immunoassays Performance Criteria for Therapeutic Drug Monitoring of Adalimumab-amgen Biosimilar (Abp501)

2020 ◽  
Author(s):  
Fabien FRANCOIS ◽  
Loubna NAIMI ◽  
Xavier ROBLIN ◽  
Anne-Emmanuelle Berger ◽  
Stephane Paul
Keyword(s):  
Human Serum ◽  
Dynamic Range ◽  
Polyclonal Antibodies ◽  
Performance Criteria ◽  
Clinical Samples ◽  
Therapeutic Drug ◽  
Serum Samples ◽  
Acceptance Criteria ◽  
Sample Stability ◽  
Validation Parameters

Abstract Background. ABP501 is a biosimilar to Reference Adalimumab (HUMIRA®) produced by AMGEN. Adalimumab (ADA) has marketing authorization for Crohn's disease, ulcerative colitis and other inflammatory or autoimmune diseases. The aim of this study was to evaluate the LISA-TRACKER assays developed by Theradiag (France), for the monitoring of ABP501 and anti-ABP501 antibodies in human serum. Methods. Accuracy of the LISA-TRACKER was measured using 3 human serum matrices spiked with known levels of biosimilar, 3 levels spanning the dynamic range. Specificity was tested with Biosimilar spiked samples, Biosimilar with Humira® spiked samples, and clinical samples from patients treated with adalimumab biosimilar. All of these samples were spiked with polyclonal antibodies directed against Humira®. Intra-run, inter-run imprecision, inhibition, kit’s stability, specificity inhibition and specificity detection steps were also part of the LISA-TRACKER Duo Adalimumab assay validation parameters. Results. 68 ABP501 clinical samples were measured with the LISA TRACKER Duo Adalimumab assay. LISA TRACKER has been validated as suitable for quantification of ABP501 in human serum samples. Concerning accuracy, percentages of recovery were ranged from 90–120% for biosimilar batch1, and between 93% and 105% for biosimilar batch2. The acceptance criteria (CV < 20%) were met for intra-run (from 3.8–16.5%) and inter-run imprecision (from 4.4–13.9%) including the two batches. All results were comprised within +/-20% from results, obtained with the kit and sample unexposed in order to evaluate stability of the sample, stability of the kit and consistency of the results. In any case but two, all percentages of inhibition were > 50% for specificity. Reagents made with ABP501 gave similar results than reagents made with Humira® meeting acceptance criteria. Conclusions. LISA-TRACKER ADA and anti-ADA assays are reliable for the monitoring of patients treated with ABP501.

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