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 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.

scholarly journals A Novel, Rapid, and Low-Volume Assay for Therapeutic Drug Monitoring of Posaconazole and Other Long-Chain Azole-Class Antifungal Drugs

mSphere ◽  
2018 ◽  
Vol 3 (6) ◽  
Author(s):  
Gregory R. Wiedman ◽  
Yanan Zhao ◽  
David S. Perlin
Keyword(s):  
Liquid Chromatography ◽  
Therapeutic Drug Monitoring ◽  
Human Serum ◽  
Drug Monitoring ◽  
Fungal Infections ◽  
Antifungal Drugs ◽  
Therapeutic Drug ◽  
Serum Samples ◽  
Human Serum Samples ◽  
Long Chain

ABSTRACT Clinicians need a better way to accurately monitor the concentration of antimicrobials in patient samples. In this report, we describe a novel, low-sample-volume method to monitor the azole-class antifungal drug posaconazole, as well as certain other long-chain azole-class antifungal drugs in human serum samples. Posaconazole represents an important target for therapeutic drug monitoring (TDM) due to its widespread use in treating invasive fungal infections and well-recognized variability of pharmacokinetics. The current “gold standard” requires trough and peak monitoring through high-pressure liquid chromatography (HPLC) or liquid chromatography-tandem mass spectroscopy (LC-MS/MS). Other methods include bioassays that use highly susceptible strains of fungi in culture plates or 96-well formats to monitor concentrations. Currently, no method exists that is both highly accurate in detecting free drug concentrations and is also rapid. Herein, we describe a new method using reduced graphene oxide (rGO) and a fluorescently labeled aptamer, which can accurately assess clinically relevant concentrations of posaconazole and other long-chain azole-class drugs in little more than 1 h in a total volume of 100 µl. IMPORTANCE This work describes an effective assay for TDM of long-chain azole-class antifungal drugs that can be used in diluted human serum samples. This assay will provide a quick, cost-effective method for monitoring concentrations of drugs such as posaconazole that exhibit well-documented pharmacokinetic variability. Our rGO-aptamer assay has the potential to improve health care for those struggling to treat fungal infections in rural or resource-limited setting.

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 P092 Development and evaluation of i-Tracker Ustekinumab and i-Tracker Anti-ustekinumab kits: fast and innovative chemiluminescent assays for the monitoring of patients treated with ustekinumab

2021 ◽  
Vol 15 (Supplement_1) ◽  
pp. S191-S191
Author(s):  
G Noguier ◽  
C Montaillier ◽  
S Daviere ◽  
Y Yang ◽  
L Colombeau ◽  
...  
Keyword(s):  
Light Emission ◽  
Inflammatory Diseases ◽  
Polyclonal Antibodies ◽  
Random Access ◽  
Therapeutic Drug ◽  
Fast Time ◽  
Serum Samples ◽  
Magnetic Microparticles ◽  
Bowel Diseases ◽  
Acridinium Ester

Abstract Background Ustekinumab, a monoclonal antibody directed against IL12/23, is a drug widely used for the treatment of Chronic Inflammatory Diseases (Psoriasis, Crohn’s disease, etc.). Therapeutic Drug Monitoring is currently proposed to provide useful information to clinicians to improve the efficacy of the treatment. Theradiag has just developed the innovative i-Tracker ustekinumab and i-Tracker Anti-ustekinumab kits: fast quantification of ustekinumab and Anti-ustekinumab antibodies fully automated on the random access i-Track10 chemiluminescent analyzer. Methods Analytical performances were assessed using 2 types of serum samples: human serum spiked with ustekinumab or Anti-ustekinumab antibodies, and samples from Inflammatory Bowel Diseases patients treated with ustekinumab (n=32). For drug measurement, ustekinumab from serum sample was captured by anti-idiotypic antibody coupled magnetic microparticles and anti-ustekinumab polyclonal antibodies conjugated to acridinium ester were used for the detection of ustekinumab. For anti-drug antibodies measurement, Anti-ustekinumab antibodies were captured according to Ustekinumab coupled magnetic microparticles and detected with the use of ustekinumab conjugated to acridinium ester. Light emission was linked to the quantity of ustekinumab, or anti-ustekinumab antibodies presents in the sample. Results Ustekinumab measurement showed high accuracy (recovery was comprised between 98% and 120%). High precision weas reached for both assays (intra-precision CV were below 9.1% and 5.3% for ustekinumab and Anti-ustekinumab assays; inter-precision CV were below 4.6% and 10.6% for ustekinumab and Anti-ustekinumab assays) and no interference was seen with biologic agents (bilirubin, hemoglobin, lipids, biotin and rheumatoid factors). The dynamic ranges of the assays were 100ng/ml to 10 000ng/ml for ustekinumab quantification and 1 AU/ml to 250 AU/ml for anti-ustekinumab antibodies quantification. i-Tracker ustekinumab and i-Tracker anti-ustekinumab assays were compared to respective ELISA based LISA-TRACKER assays and showed excellent correlation (R² = 0.96, Slope = 0.93 for ustekinumab assay; Spearman’s coefficient correlation was 0.95 for Anti-ustekinumab assay). Conclusion i-Tracker kits are innovative assays which exhibit fast (time to results &lt; 40min), accurate and reproducible results for the quantification of ustekinumab and Anti-ustekinumab antibodies. Excellent agreements were observed with respective LISA-TRACKER assays. i-Tracker kits are valuable tools for the monitoring of patients treated with ustekinumab.

scholarly journals Reusable, Noninvasive, and Sensitive Fluorescence Enhanced ZnO-Nanorod-Based Microarrays for Quantitative Detection of AFP in Human Serum

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Saima Rafique ◽  
Farukh Kiyani ◽  
Sumbal Jawaid ◽  
Rubina Nasir ◽  
Mahmoosh Ahmad ◽  
...  
Keyword(s):  
Human Serum ◽  
Dynamic Range ◽  
Limit Of Detection ◽  
Zno Nanorods ◽  
Protein Microarrays ◽  
Great Promise ◽  
Quantitative Detection ◽  
Zno Nanorod ◽  
Serum Samples ◽  
Protein Assay

The fabrication of sensitive protein microarrays such as PCR used in DNA microarray is challenging due to lack of signal amplification. The development of microarrays is utilized to improve the sensitivity and limitations of detection towards primal cancer detection. The sensitivity is enhanced by the use of ZnO-nanorods and is investigated as a substrate which enhance the florescent signal to diagnose the hepatocellular carcinoma (HCC) at early stages. The substrate for deposition of ZnO-nanorods is prepared by the conventional chemical bath deposition method. The resultant highly dense ZnO-nanorods enhance the fluorescent signal 7.2 times as compared to the substrate without ZnO-nanorods. The microarray showed sensitivity of 1504.7 ng ml-1 and limit of detection of 0.1 pg ml-1 in wide dynamic range of 0.05 pg-10 μg ml-1 for alpha fetoprotein (AFP) detection in 10% human serum. This immunoassay was successfully applied for human serum samples to detect tumor marker with good recoveries. The ZnO-nanorod substrate is a simple protein microarray which showed a great promise for developing a low-cost, sensitive, and high-throughput protein assay platform for several applications in both fundamental research and clinical diagnosis.

scholarly journals Development of an UPLC/MS–MS method for quantification of intact IGF-I from human serum

Bioanalysis ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 53-65
Author(s):  
Nikunj N Tanna ◽  
Mary E Lame ◽  
Mark Wrona
Keyword(s):  
Protein Binding ◽  
Human Serum ◽  
Dynamic Range ◽  
Multiple Reaction Monitoring ◽  
Sodium Dodecyl ◽  
Elevated Serum ◽  
Analytical Sensitivity ◽  
Solid Core ◽  
Serum Samples ◽  
Igf I

Aim: Developing LC–MS methods for biomolecules is often challenging due to issues with molecular size and complexity, nonspecific binding, protein binding, solubility and sensitivity. As a result, complex sample preparation workflows, including immune-affinity and/or protein digestion and lengthy analysis potentially using nano-flow LC, may be needed to achieve the required sensitivity. This work aims to provide a simple, sensitive, fast and robust method for quantification of intact IGF-I from human serum using UPLC–MS/MS. Methods: IGF-I serum samples were denatured with sodium dodecyl sulfate, followed by organic protein precipitation to effectively disrupt protein binding and subsequent SPE of the resulting supernatant for sample cleanup and enrichment prior to LC–MS/MS analysis. Separation was performed on an analytical scale LC using a reversed-phase column containing <2 μm solid core particle followed by detection on a tandem quadrupole MS in multiple reaction monitoring mode. Results: Intact IGF-I was quantified from serum using the method described above at a LLOQ of 5 ng/ml with a dynamic range 5–1000 ng/ml (r2>0.99) and mean accuracy of 101.76%. Accuracies for quality control samples were between 93.9–107.7% with RSD <7%. Conclusion: The analytical sensitivity, linear dynamic range and excellent reproducibility of this method reliably measures endogenous and elevated serum IGF-I levels, demonstrating its utility in discovery, bioanalysis and clinical research.

Role of rapid immunoassays for urgent ("stat") determinations of creatine kinase isoenyme MB.

1989 ◽  
Vol 35 (8) ◽  
pp. 1752-1756 ◽  
Author(s):  
A H Wu ◽  
T G Gornet ◽  
C C Harker ◽  
H L Chen
Keyword(s):  
Myocardial Infarction ◽  
Creatine Kinase ◽  
Early Diagnosis ◽  
Artery Bypass ◽  
Performance Criteria ◽  
Analytical Sensitivity ◽  
Serum Samples ◽  
Assay Performance ◽  
Sample Stability

Abstract We compared the analytical performance of three immunoassays used to rapidly determine creatine kinase (EC 2.7.3.2; CK) isoenzyme MB in serum: Dade's "Stratus," Corning's "Magic Lite," and Hybritech's "Icon QSR CK-MB." Performance criteria included precision, analytical sensitivity, sample stability, and analytical and clinical correlation of results for serum samples taken from healthy individuals, patients with suspected and confirmed acute myocardial infarction, and patients after coronary artery bypass surgery. We also examined 31 samples taken from patients in the emergency room suspected of myocardial infarction, to evaluate the potential of these assays for early diagnosis. Although these assays differ in the manner in which CK-MB is measured, and therefore have different procedural requirements, we conclude that they are equivalent in overall assay performance. None of these assays, however, is sufficiently sensitive for early diagnosis of myocardial infarction; therefore, results cannot be used by cardiologists in deciding whether acute thrombolytic therapy should be given. Other management decisions, such as the optimal utilization of intensive-care bed space, may justify using these assays on a "stat" basis.

Measurement of mycophenolic acid in plasma or serum by a commercial enzyme inhibition technique in comparison with a high performance liquid chromatography method

2008 ◽  
Vol 46 (9) ◽  
Author(s):  
Dustin R. Bunch ◽  
Sihe Wang
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Enzyme Inhibition ◽  
Mycophenolic Acid ◽  
High Performance ◽  
Random Access ◽  
Hplc Method ◽  
Clinical Samples ◽  
Therapeutic Drug ◽  
Serum Samples

Abstract: Mycophenolic acid (MPA) is the primary active metabolite of the immunosuppressant mycophenolate mofetil. High performance liquid chromatography (HPLC) is commonly used for therapeutic drug monitoring (TDM) of MPA but requires batched runs. Recently, an enzyme inhibition assay (EIA) was approved for MPA TDM on random-access platforms using either serum or EDTA plasma. We evaluated the EIA on a Roche Integra 400 using serum and heparinized plasma in comparison with a validated HPLC method.: Heparinized plasma from leftover clinical samples on which MPA was ordered along with paired serum samples, drawn at the same time for other clinical tests, were used for the method comparison.: The EIA was linear from 3.1 to 44.0 μmol/L with an accuracy of 93.9%–107.1%. The intra- and inter-day variations were 0.5%–2.7% and 1.6%–2.1%, respectively. The limit of detection was 0.8 μmol/L and the limit of quantification was 3.1 μmol/L. The method showed a mean bias of 0.6 μmol/L (7.6%) in serum samples (3.1–34.1 μmol/L) vs. the HPLC method using paired plasma (n=229). Heparinized plasma (n=114) vs. serum showed a mean bias of –0.1 μmol/L (–1.6%) by the EIA.: The random-access EIA on Integra 400 is acceptable for clinical MPA TDM in either serum or heparinized plasma.Clin Chem Lab Med 2008;46:1281–4.

scholarly journals Differentiating iron-loading anemias using a newly developed and analytically validated ELISA for human serum erythroferrone

PLoS ONE ◽  
2021 ◽  
Vol 16 (7) ◽  
pp. e0254851
Author(s):  
Laura Diepeveen ◽  
Rian Roelofs ◽  
Nicolai Grebenchtchikov ◽  
Rachel van Swelm ◽  
Leon Kautz ◽  
...  
Keyword(s):  
Human Serum ◽  
Polyclonal Antibodies ◽  
Sandwich Elisa ◽  
Thalassemia Major ◽  
Diagnostic Value ◽  
Iron Availability ◽  
Analytical Validation ◽  
Serum Samples ◽  
Reliable Quantification ◽  
First Time

Erythroferrone (ERFE), the erythroid regulator of iron metabolism, inhibits hepcidin to increase iron availability for erythropoiesis. ERFE plays a pathological role during ineffective erythropoiesis as occurs in X-linked sideroblastic anemia (XLSA) and β-thalassemia. Its measurement might serve as an indicator of severity for these diseases. However, for reliable quantification of ERFE analytical characterization is indispensable to determine the assay’s limitations and define proper methodology. We developed a sandwich ELISA for human serum ERFE using polyclonal antibodies and report its extensive analytical validation. This new assay showed, for the first time, the differentiation of XLSA and β-thalassemia major patients from healthy controls (p = 0.03) and from each other (p<0.01), showing the assay provides biological plausible results. Despite poor dilution linearity, parallelism and recovery in patient serum matrix, which indicated presence of a matrix effect and/or different immunoreactivity of the antibodies to the recombinant standard and the endogenous analyte, our assay correlated well with two other existing ERFE ELISAs (both R2 = 0.83). Nevertheless, employment of one optimal dilution of all serum samples is warranted to obtain reliable results. When adequately performed, the assay can be used to further unravel the human erythropoiesis-hepcidin-iron axis in various disorders and assess the added diagnostic value of ERFE.

An UHPLC-UV Method for Determination of Vancomycin in human serum

2020 ◽  
Vol 16 ◽  
Author(s):  
Fang Fang ◽  
Ning Li ◽  
Chunli Xu ◽  
Rong Tan ◽  
Jihong Yang ◽  
...  
Keyword(s):  
Therapeutic Drug Monitoring ◽  
Human Serum ◽  
Drug Monitoring ◽  
Internal Standard ◽  
Sample Pretreatment ◽  
Gradient Elution ◽  
Therapeutic Drug ◽  
Serum Samples ◽  
Column Temperature ◽  
Limit Of Quantitation

Objective: To develop a rapid ultra-performance liquid chromatographic (UHPLC)-UV method for vancomycin determination in human serum for therapeutic drug monitoring (TDM). Methods: Human serum samples were precipitated with 10% perchloric acid, and the supernatant after centrifugation was analyzed on an ACQUITY UHPLC BEH C18 column (2.1 × 50mm, 1.7 μm) via gradient elution with a flow rate at 0.3 mL/min. The mobile phase consisted of acetonitrile and 0.005M KH2PO4 buffer (containing 0.1% triethylamine, pH 3.4). The detection wavelength was set at 210 nm, and the column temperature was set at 40. The total runtime was 6.0 min per analysis. Results: After comprehensive validation, the method was applied to determine the concentration of vancomycin in human serum. The chromatographic peaks of vancomycin and internal standard were not interfered by endogenous matrixes. The retention time (RT) of vancomycin was 1.91 min, while the internal standard was 1.58 min. The good linearity range of vancomycin concentration was 2.5-120 μg/mL (R2>0.999). The lower limit of quantitation (LLOQ) was 2.5 μg/mL. The precision at three quality control (QC) levels (including LLOQ) was restricted within 85-115%. The extraction recovery rate of QC samples (4.0, 20.0, 60.0 μg/mL) were 101.16%、97.70%、94.90%, respectively. Inter- and intra-day precision was less than 8% (RSD). Stability tests under different storage conditions were satisfactory. In patients, the concentration of vancomycin ranged from 7.30 to 89.12 μg/mL determined by the fully validated method. Conclusion: The simple, rapid sample pretreatment procedures and short analysis time made this UHPLC-UV method suitable for therapeutic drug monitoring (TDM) of vancomycin.

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