Therapeutic Drug Monitoring: Performance of a FRET-Based Point-Of-Care Immunoassay for the Quantitation of Infliximab and Adalimumab in Blood

Two fast (<5 min), time-resolved fluorescence resonance energy transfer (FRET)-based immunoassays (Procise IFX™ and Procise ADL™) were developed for the quantitative detection of infliximab (IFX), adalimumab (ADL), and their respective biosimilars for use in therapeutic drug monitoring (TDM) using 20 µL of finger prick whole blood at the point-of-care or whole blood/serum in a central lab. Studies were performed to characterize analytical performance of the Procise IFX and the Procise ADL assays on the ProciseDx™ analyzer. The Procise IFX and Procise ADL assays both showed good analytical performance with respect to sensitivity, specificity, linearity, and precision suitable for routine clinical use as well as excellent correlation to current commercial ELISA IFX and ADL measurement methods. Results indicated that the Procise IFX and Procise ADL assays are sensitive, specific, and precise yielding results in less than 5 minutes from either whole blood or serum. This indicates the Procise IFX and Procise ADL assays are useful for obtaining fast and accurate IFX or ADL quantitation, thus avoiding delays inherent to current methods and enabling immediate drug level dosing decisions to be made during a single patient visit.

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Therapeutic Drug Monitoring: Performance of a FRET-Based Point-Of-Care Immunoassay for the Quantitation of Infliximab and Adalimumab in Blood

10.26434/chemrxiv.13277321.v2 ◽

2020 ◽

Author(s):

Edgar Ong ◽

Ruo Huang ◽

Richard Kirkland ◽

Stefan Westin ◽

Jared Salbato ◽

...

Keyword(s):

Therapeutic Drug Monitoring ◽

Drug Monitoring ◽

Whole Blood ◽

Point Of Care ◽

Resonance Energy Transfer ◽

Resonance Energy ◽

Drug Level ◽

Analytical Performance ◽

Quantitative Detection ◽

Therapeutic Drug

Two fast (<5 min), time-resolved fluorescence resonance energy transfer (FRET)-based immunoassays (Procise IFX™ and Procise ADL™) were developed for the quantitative detection of infliximab (IFX), adalimumab (ADL), and their respective biosimilars for use in therapeutic drug monitoring (TDM) using 20 µL of finger prick whole blood at the point-of-care or whole blood/serum in a central lab. Studies were performed to characterize analytical performance of the Procise IFX and the Procise ADL assays on the ProciseDx™ analyzer. The Procise IFX and Procise ADL assays both showed good analytical performance with respect to sensitivity, specificity, linearity, and precision suitable for routine clinical use as well as excellent correlation to current commercial ELISA IFX and ADL measurement methods. Results indicated that the Procise IFX and Procise ADL assays are sensitive, specific, and precise yielding results in less than 5 minutes from either whole blood or serum. This indicates the Procise IFX and Procise ADL assays are useful for obtaining fast and accurate IFX or ADL quantitation, thus avoiding delays inherent to current methods and enabling immediate drug level dosing decisions to be made during a single patient visit.

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Rapid analysis of whole blood by paper spray mass spectrometry for point-of-care therapeutic drug monitoring

The Analyst ◽

10.1039/c2an35082c ◽

2012 ◽

Vol 137 (10) ◽

pp. 2344 ◽

Cited By ~ 112

Author(s):

Ryan D. Espy ◽

Nicholas E. Manicke ◽

Zheng Ouyang ◽

R. Graham Cooks

Keyword(s):

Mass Spectrometry ◽

Therapeutic Drug Monitoring ◽

Drug Monitoring ◽

Whole Blood ◽

Point Of Care ◽

Rapid Analysis ◽

Therapeutic Drug ◽

Paper Spray

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P384 Therapeutic drug monitoring: performance of the first lateral flow-based Point of Care test for the quantification of infliximab in a finger prick

Journal of Crohn s and Colitis ◽

10.1093/ecco-jcc/jjab076.508 ◽

2021 ◽

Vol 15 (Supplement_1) ◽

pp. S396-S397

Author(s):

A Ametzazurra ◽

J Pascual ◽

L Del Rio ◽

A Maguregui ◽

D Nagore ◽

...

Keyword(s):

Therapeutic Drug Monitoring ◽

Drug Monitoring ◽

Whole Blood ◽

Limit Of Detection ◽

Point Of Care ◽

Package Insert ◽

Lateral Flow ◽

Therapeutic Drug ◽

Limit Of Quantification ◽

Finger Prick

Abstract Background Promonitor Quick IFX is a lateral flow test (LFT) for the quantification of infliximab (IFX) in human whole blood (finger prick or venous) or serum in 20 minutes. This LFT is based on a sandwich immunoassay to quantify either the reference IFX or biosimilars. This abstract describes the studies performed to establish the analytical specifications of the product. Methods Clinical and Laboratory Standards Institute (CLSI) guidelines were followed for the evaluation of the analytical specifications of the LFT in whole blood and serum matrices: Linearity (EP-06-A), Detection capability (EP17-A2), Interfering substances (EP07, 3rd Edition), Intermediate precision (EP05-A3) and Bias evaluation study for Biosimilars (EP10-A3). Results were obtained in combination with the automated portable reader PQreader. A Datamatrix provided with each Promonitor Quick IFX kit contains the calibration information required for the PQreader to measure the Control and Test lines and report the IFX concentration. Results The linear assay range was determined to be 1-58 µg/mL in whole-blood and 0.6-67 µg/mL in serum according to the processes indicated in the Package Insert. The Limit of Blank is 0.8 μg/mL, the Limit of Detection and Lower Limit of Quantification (LLoQ) are 1.1 μg/mL, and the Upper Limit of Quantification (ULoQ) is 15.4 μg/mL. There was no effect on assay performance when each of the following substances were added to samples with 0, 3, and 7 μg/mL of IFX: Haemoglobin (>1000 mg/dL), Bilirubin (>40 mg/dL), Triglycerides (>1500 mg/dL), HAMA (160 AU/mL), Rheumatoid factor (200 IU/mL), EDTA (5.4 mg/mL), Heparin (51 U/mL), Citrate (11.4%), Vedolizumab (60 μg/mL) and Adalimumab (20.25 μg/mL). Repeatability and within-device precision results obtained for the positive samples are shown in the table below. The negative samples showed a negative result in all the measurements. A bias study showed that Promonitor Quick IFX can quantify CT-P13, SB2 and GP1111 biosimilars throughout the measurement range with a maximum bias of 14%. Conclusion Promonitor Quick IFX is the first LFT available for true Point of Care testing of patients treated with IFX with just a finger prick sample. It provides quick turnaround time to facilitate therapeutic drug monitoring and aid immediate decision making in the doctor office or hospitals with an excellent analytical performance.

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Performance of a FRET-Based Point-of-Care Immunoassay for the Quantitation of Fecal Calprotectin

10.26434/chemrxiv.14043821.v1 ◽

2021 ◽

Author(s):

Manisha Yadav ◽

Michael Skinner ◽

Rukmini Reddy ◽

Matthew Wong ◽

Kevin Chon ◽

...

Keyword(s):

Point Of Care ◽

Resonance Energy Transfer ◽

Resonance Energy ◽

Fecal Calprotectin ◽

Analytical Performance ◽

Quantitative Detection ◽

Time Resolved ◽

Collection Device ◽

Sensitivity Specificity ◽

A Current

A fast (~5 min), time-resolved fluorescence resonance energy transfer based immunoassay (Procise FCP<a>™</a>) was developed for the point-of-care quantitative detection of fecal calprotectin (FCP) using 15 mg of fecal specimen eluted in collection fluid from the Procise Stool Collection Device™. Studies were performed to characterize analytical performance of the Procise FCP assay on the ProciseDx™ analyzer. The Procise FCP assay showed good analytical performance with respect to sensitivity, specificity, linearity, and precision suitable for routine clinical use in a point-of-care setting as well as excellent analytical agreement with a current commercial FCP measurement method. Results indicate that the Procise FCP assay is sensitive, specific, and precise yielding results in less than 5 minutes. This indicates the Procise FCP assay is useful for obtaining fast and accurate FCP quantitation, thus avoiding delays inherent to current methods and enabling immediate clinical assessment to be made during a single patient visit.

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Performance of a FRET-Based Point-of-Care Immunoassay for the Quantitation of Fecal Calprotectin

10.26434/chemrxiv.14043821 ◽

2021 ◽

Author(s):

Manisha Yadav ◽

Michael Skinner ◽

Rukmini Reddy ◽

Matthew Wong ◽

Kevin Chon ◽

...

Keyword(s):

Point Of Care ◽

Resonance Energy Transfer ◽

Resonance Energy ◽

Fecal Calprotectin ◽

Analytical Performance ◽

Quantitative Detection ◽

Time Resolved ◽

Collection Device ◽

Sensitivity Specificity ◽

A Current

A fast (~5 min), time-resolved fluorescence resonance energy transfer based immunoassay (Procise FCP<a>™</a>) was developed for the point-of-care quantitative detection of fecal calprotectin (FCP) using 15 mg of fecal specimen eluted in collection fluid from the Procise Stool Collection Device™. Studies were performed to characterize analytical performance of the Procise FCP assay on the ProciseDx™ analyzer. The Procise FCP assay showed good analytical performance with respect to sensitivity, specificity, linearity, and precision suitable for routine clinical use in a point-of-care setting as well as excellent analytical agreement with a current commercial FCP measurement method. Results indicate that the Procise FCP assay is sensitive, specific, and precise yielding results in less than 5 minutes. This indicates the Procise FCP assay is useful for obtaining fast and accurate FCP quantitation, thus avoiding delays inherent to current methods and enabling immediate clinical assessment to be made during a single patient visit.

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PERFORMANCE CHARACTERISTICS OF A FRET-BASED IMMUNOASSAY FOR QUANTITATION OF INFLIXIMAB ON A POINT-OF-CARE INSTRUMENT SYSTEM

Inflammatory Bowel Diseases ◽

10.1093/ibd/izaa347.135 ◽

2021 ◽

Vol 27 (Supplement_1) ◽

pp. S57-S57

Author(s):

Edgar Ong ◽

Ruo Huang ◽

Richard Kirkland ◽

Michael Hale ◽

Larry Mimms

Keyword(s):

Whole Blood ◽

Limit Of Detection ◽

Point Of Care ◽

Quantitative Detection ◽

Therapeutic Drug ◽

Analytical Sensitivity ◽

Sample Type ◽

Hook Effect ◽

Limit Of Quantitation ◽

Assay Precision

Abstract Introduction A fast (<5 min), time-resolved fluorescence resonance energy transfer (FRET)-based immunoassay was developed for the quantitative detection of infliximab (IFX) and biosimilars for use in therapeutic drug monitoring using only 20 µL of fingerstick whole blood or serum at the point-of-care. The Procise IFX assay and ProciseDx analyzer are CE-marked. Studies were performed to characterize analytical performance of the Procise IFX assay on the ProciseDx analyzer. Methods Analytical testing was performed by spiking known amounts of IFX into negative serum and whole blood specimens. Analytical sensitivity was determined using limiting concentrations of IFX. Linearity was determined by testing IFX across the assay range. Hook effect was assessed at IFX concentrations beyond levels expected to be found within a patient. Testing of assay precision, cross-reactivity and potential interfering substances, and biosimilars was performed. The Procise IFX assay was also compared head-to-head with another CE-marked assay: LISA-TRACKER infliximab ELISA test (Theradiag, France). The accuracy of the Procise IFX assay is established through calibrators and controls traceable to the WHO 1st International Standard for Infliximab (NIBSC code: 16/170). Results The Procise IFX assay shows a Limit of Blank, Limit of Detection, and Lower Limit of Quantitation (LLoQ) of 0.1, 0.2, and 1.1 µg/mL in serum and 0.6, 1.1, and 1.7 µg/mL in whole blood, respectively. The linear assay range was determined to be 1.7 to 77.2 µg/mL in serum and whole blood. No hook effect was observed at an IFX concentration of 200 µg/mL as the value reported as “>ULoQ”. Assay precision testing across 20 days with multiple runs and reagent lots showed an intra-assay coefficient of variation (CV) of 2.7%, an inter-assay CV of <2%, and a total CV of 3.4%. The presence of potentially interfering/cross-reacting substances showed minimal impact on assay specificity with %bias within ±8% of control. Testing of biosimilars (infliximab-dyyb and infliximab-abda) showed good recovery. A good correlation to the Theradiag infliximab ELISA was obtained for both serum (slope=1.01; r=0.99) and whole blood (slope=1.01; r=0.98) samples (Figure 1). Conclusion Results indicate that the Procise IFX assay is sensitive, specific, and precise yielding results within 5 minutes from both whole blood and serum without the operator needing to specify sample type. Additionally, it shows very good correlation to a comparator assay that takes several hours and sample manipulation to yield results. This makes the Procise IFX assay ideal for obtaining fast and accurate IFX quantitation, thus allowing for immediate drug level dosing decisions to be made by the physician during patient treatment.

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Rapid point-of-care anti-infliximab antibodies detection in clinical practice: comparison with ELISA and potential for improving therapeutic drug monitoring in IBD patients

Therapeutic Advances in Gastroenterology ◽

10.1177/1756284821999902 ◽

2021 ◽

Vol 14 ◽

pp. 175628482199990

Author(s):

Sonia Facchin ◽

Andrea Buda ◽

Romilda Cardin ◽

Nada Agbariah ◽

Fabiana Zingone ◽

...

Keyword(s):

Clinical Practice ◽

Therapeutic Drug Monitoring ◽

Drug Monitoring ◽

Point Of Care ◽

Drug Clearance ◽

Elisa Test ◽

Enzyme Linked Immunosorbent Assay ◽

Therapeutic Drug ◽

Drug Concentrations ◽

Inflammatory Bowel

Anti-drug antibodies can interfere with the activity of anti-tumor necrosis factor (TNF) agents by increasing drug clearance via direct neutralization. The presence of anti-drug antibodies is clinically relevant when trough drug concentrations are undetectable or sub-therapeutic. However, traditional immunoassay is not easily and rapidly accessible, making the translation of the results into treatment adjustment difficult. The availability of a point-of-care (POC) test for therapeutic drug monitoring (TDM) might represent an important step forward for improving the management of inflammatory bowel disease (IBD) patients in clinical practice. In this pilot study, we compared the results obtained with POC tests with those obtained by enzyme-linked immunosorbent assay (ELISA) in a group of IBD patients treated with Infliximab (IFX). We showed that POC test can reliably detect presence of antibody-to-IFX with 100% of specificity and 76% sensitivity, in strong agreement with the ELISA test ( k-coefficient = 0.84).

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P465 Therapeutic drug monitoring in Crohn’s disease patients, a comparison between homogeneous mobility shift assay and point of care method

Journal of Crohn s and Colitis ◽

10.1093/ecco-jcc/jjz203.594 ◽

2020 ◽

Vol 14 (Supplement_1) ◽

pp. S412-S412

Author(s):

G Bodini ◽

M G Demarzo ◽

A Djahandideh ◽

I Baldissarro ◽

E Savarino ◽

...

Keyword(s):

Crohn’S Disease ◽

Therapeutic Drug Monitoring ◽

Crohn's Disease ◽

Drug Concentration ◽

Drug Monitoring ◽

Point Of Care ◽

Therapeutic Drug ◽

Serum Drug Concentration ◽

Disease Relapse

Abstract Background Therapeutic Drug Monitoring (TDM) is a useful tool to help physicians managing patients with Inflammatory Bowel Disease treated with anti-tumour necrosis factor (TNF) drugs. Different techniques are available to evaluate serum drug concentration (TL), However, these techniques are time-consuming. A point-of-care (POC) method has been proposed to evaluate drug TL and overcome the limitations inherent to other methodologies. Our aim was to evaluate the capability of POC to discriminate between IBD relapse and remission and to evaluate the concordance of drug TL measured with POC and HMSA Methods We analysed with Quantum BlueÒ (Buhlmann Laboratories AG, Schonenbuch, Switzerland) (POC) 200 Adalimumab (ADA) and 200 Infliximab serum samples of 46 Crohn’s disease (CD) patients previously assessed with HMSA. Blood samples were drawn at standardised time points during anti-TNF treatment (2, 6, and every 8 weeks), before anti-TNF administration. Disease activity was assessed by the Harvey–Bradshaw Index (HBI, remission defined by HBI<5). Results We evaluated 46 CD patients responders to anti-TNF induction with ADA (n = 25, 54.3%) and IFX (n = 21, 45.6%) with a median follow-up of 83 weeks (range 16–144 weeks). At week 16, median ADA TL of patients in remission were significantly higher as compared with patients in disease relapse using both HMSA [12.7 μg/ml (range, 8.9–23.6 μg/ml) vs. 6.6 μg/ml (range, 0.7–9.6 μg/ml), p = 0.0001] and POC [17.8 μg/ml (range 7.6–35.0 μg/ml) vs. 9.8 μg/ml (range 5.8–11.4 μg/ml), p = 0.0003]. The concordance between the two different techniques has been assessed as 0.76 by Choen Kappa. Considering IFX TL, patients in remission had higher serum drug concentration using both HMSA [7.0 μg/ml (range, 0.0–21.8 μg/ml)] and POC [6.2 μg/ml (range 0.4–14.3 μg/ml)] as compared with patients who experienced disease relapse [HMSA, 0.1 μg/ml (range, 0.0–4.1 μg/ml), p = 0.019; POC, 0.45 μg/ml (range 0.4–3.3 μg/ml), p = 0.0072]. The concordance between the two different test for IFX TL was 0.81. We obtained similar results at the end of follow-up: median ADA TL was higher in remission than in disease relapse patients using both HMSA and POC [p = 0.001 and p = 0.0012] with a concordance of 0.75. Median IFX TL was higher in remission than in disease relapse patients using both HMSA and POC (p = 0.13 and p = 0.25) with a concordance of 0.70. Conclusion Both POC and HMSA are TL tests able to differentiate relapse and remission in IBD patients. The association between anti-TNF TL and disease status (remission/relapse) was better in ADA-treated patients rather than patients treated with IFX. Finally, we demonstrated a good concordance between HMSA and POC. Anti-drug antibody concentrations while available on HMSA were not available on POC

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