Abstract 4541: Highly sensitive kinase assays combining Carna Biosciences QSS Assist ELISA reagents with the Meso Scale Discovery MULTI-ARRAY platform.

Background: Anti-drug antibodies (ADA) against natalizumab develop early during treatment. ADA persistency is defined by two consecutive positive results as performed by the current qualitative ELISA assay (positive/negative). Very little is known about the magnitude of the natalizumab ADA response and persistency. Design/methods: We developed a highly sensitive natalizumab ADA titration assay on the Meso Scale Discovery (MSD) platform and a pharmacokinetic (PK) assay. We included 43 patients with a positive ELISA-ADA result within 6 months of treatment initiation (baseline) of whom a follow-up serum sample was available 12–30 months after treatment start. MSD-ADA titres and drug levels were measured. Results: Median MSD-ADA titre at baseline was 4881 and 303 at follow-up. A titre of >400 at baseline had a 94% sensitivity and 89% specificity to predict ADA persistency. Reversion to ADA negativity occurred in 10 patients with mean drug levels of 10.8 μg/mL. The median trough drug level in ADA-positive samples was 0 µg/mL. PK levels and ADA titres correlated strongly negatively ( r = −0.67). Conclusion: High baseline natalizumab ADA titres accurately predict persistency. Despite continuous treatment, the majority of patients with persistent ADA had no detectable drug levels indicating loss of efficacy in line with phase 3 study results.

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Emerging Technologies and Generic Assays for the Detection of Anti-Drug Antibodies

Journal of Immunology Research ◽

10.1155/2016/6262383 ◽

2016 ◽

Vol 2016 ◽

pp. 1-6 ◽

Cited By ~ 15

Author(s):

Michael A. Partridge ◽

Shobha Purushothama ◽

Chinnasamy Elango ◽

Yanmei Lu

Keyword(s):

Patient Safety ◽

Surface Plasmon Resonance ◽

Drug Development ◽

Surface Plasmon ◽

Plasmon Resonance ◽

Assay Development ◽

Meso Scale Discovery ◽

Antibody Assays ◽

Drug Pharmacokinetics ◽

Meso Scale

Anti-drug antibodies induced by biologic therapeutics often impact drug pharmacokinetics, pharmacodynamics response, clinical efficacy, and patient safety. It is critical to assess the immunogenicity risk of potential biotherapeutics in producing neutralizing and nonneutralizing anti-drug antibodies, especially in clinical phases of drug development. Different assay methodologies have been used to detect all anti-drug antibodies, including ELISA, radioimmunoassay, surface plasmon resonance, and electrochemiluminescence-based technologies. The most commonly used method is a bridging assay, performed in an ELISA or on the Meso Scale Discovery platform. In this report, we aim to review the emerging new assay technologies that can complement or address challenges associated with the bridging assay format in screening and confirmation of ADAs. We also summarize generic anti-drug antibody assays that do not require drug-specific reagents for nonclinical studies. These generic assays significantly reduce assay development efforts and, therefore, shorten the assay readiness timeline.

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A competitive ligand-binding assay to detect neutralizing antibodies to a bispecific drug using a used multiplex Meso Scale Discovery platform

Bioanalysis ◽

10.4155/bio-2021-0177 ◽

2021 ◽

Author(s):

Vitaly Ablamunits ◽

Soma Basak ◽

Rosemary Lawrence-Henderson ◽

Teresa M Caiazzo ◽

John Kamerud

Keyword(s):

Ligand Binding ◽

Neutralizing Antibodies ◽

Binding Assay ◽

Large Molecule ◽

Ligand Binding Assay ◽

Meso Scale Discovery ◽

Candidate Drug ◽

Meso Scale ◽

Competitive Ligand ◽

Better Than

Background: Monitoring appearance of neutralizing antibodies (NAbs) to multidomain large molecule drugs is a challenging task. Materials & methods: Here, we report development of a competitive ligand-binding assay for detection of NAbs to a bispecific candidate drug using a used multiplex Meso Scale Discovery platform, which allows for detection of NAbs to both drug arms in the same sample. Results: The assay has sensitivity better than 250 ng/ml and is tolerant to the presence of drug at concentration >600 μg/ml and to the level of soluble target(s) >400 ng/ml. Conclusion: Our data suggest that multiplex approach can be successfully used for development of NAb assays in competitive ligand-binding assay format.

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Multiplex measurement of proinflammatory cytokines in human serum: Comparison of the Meso Scale Discovery electrochemiluminescence assay and the Cytometric Bead Array

Journal of Immunological Methods ◽

10.1016/j.jim.2011.06.033 ◽

2011 ◽

Vol 372 (1-2) ◽

pp. 71-77 ◽

Cited By ~ 70

Author(s):

Djeneba Dabitao ◽

Joseph B. Margolick ◽

Joseph Lopez ◽

Jay H. Bream

Keyword(s):

Human Serum ◽

Proinflammatory Cytokines ◽

Cytometric Bead Array ◽

Meso Scale Discovery ◽

Meso Scale

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Use of Meso-Scale Discovery™ to Examine Cytokine Content in Microglia Cell Supernatant

Microglia - Methods in Molecular Biology ◽

10.1007/978-1-62703-520-0_11 ◽

2013 ◽

pp. 93-100 ◽

Cited By ~ 9

Author(s):

Miguel A. Burguillos

Keyword(s):

Meso Scale Discovery ◽

Microglia Cell ◽

Meso Scale ◽

Cell Supernatant

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Meso Scale Discovery and Luminex Comparative Analysis of Calbindin D28K

Journal of Biomedicine and Biotechnology ◽

10.1155/2009/187426 ◽

2009 ◽

Vol 2009 ◽

pp. 1-5 ◽

Cited By ~ 6

Author(s):

Samer Sourial ◽

Maritha Marcusson-Ståhl ◽

Karin Cederbrant

Keyword(s):

Comparative Analysis ◽

Human Kidney ◽

Distal Tubule ◽

Specific Protein ◽

Tubular Damage ◽

Meso Scale Discovery ◽

Multiplex Immunoassay ◽

Pathological Conditions ◽

Calbindin D28k ◽

Meso Scale

The sensitivity of different renal regions to xenobiotics requires the development of a multiplex immunoassay for the simultaneous analysis of kidney biomarkers. Calbindin D28K is a distal tubule-specific protein that can be detected in urine under pathological conditions. In this study, a pair of anti-calbindin D28K antibodies was used in an immunoassay for the detection of calbindin D28K expression in rat and human kidney and urine. Comparative analysis of the immunoassay was performed on the Meso Scale Development (MSD) and Luminex platforms. Analysis on both platforms detected calbindin D28K concentrations between 100 ng/mL and 100 pg/mL. Luminex detected 10-fold the amount of calbindin D28K in samples analyzed as compared to MSD, whereas calbindin D28K level in rat and human urine was below detection limit in both platforms. The application of the immunoassays described herein may be useful in toxicological and pathological studies of distal tubular damage in rats and human.

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