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

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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Optimization and Validation of a Multiplex, Electrochemiluminescence-Based Detection Assay for the Quantitation of Immunoglobulin G Serotype-Specific Antipneumococcal Antibodies in Human Serum

Clinical and Vaccine Immunology ◽

10.1128/cvi.00415-08 ◽

2009 ◽

Vol 16 (3) ◽

pp. 387-396 ◽

Cited By ~ 55

Author(s):

Rocio D. Marchese ◽

Derek Puchalski ◽

Pamela Miller ◽

Joseph Antonello ◽

Olivia Hammond ◽

...

Keyword(s):

Human Serum ◽

Immunoglobulin G ◽

Dynamic Range ◽

Cross Reactivity ◽

Enzyme Linked Immunosorbent Assay ◽

Testing Time ◽

Igg Antibody ◽

Meso Scale Discovery ◽

Acceptance Criterion ◽

Detection Assay

ABSTRACT Pneumovax 23 consists of a mixture of highly purified capsular polysaccharides (Ps) from 23 of the most prevalent serotypes of Streptococcus pneumoniae. Testing of vaccine immunogenicity has been historically performed on the enzyme-linked immunosorbent assay (ELISA) platform, validated to measure immunoglobulin G (IgG) antibodies to all 23 serotypes included in Pneumovax 23. In order to significantly improve the throughput of this form of testing, we have developed and validated a direct binding electrochemiluminescence (ECL)-based multiplex assay that can measure the antibody response in human serum to eight serotypes within a single microtiter well. The pneumococcal (Pn) ECL assay is based on the Meso Scale Discovery (MSD) technology which utilizes a Sulfo-Tag-labeled anti-human IgG antibody that emits light upon electrochemical stimulation. The Pn ECL assay exhibits a wide dynamic range and provides the ability to read concentrations down to the minimum reported concentration in the Merck ELISA (0.1 μg/ml). Cross-reactivity assessment using type-specific monoclonal antibodies showed no cross talk between antigen spots within a well. By use of the WHO Pn sample reference panel, the results obtained with the Pn ECL assay were compared to the results obtained with the international Pn ELISA. The results for the Pn ECL assay satisfied the WHO-recommended acceptance criterion for concordance for all seven serotypes with published Pn ELISA values, and the overall correlation (r value) across the seven serotypes was 0.994. The MSD methodology has great potential to be extremely useful for simultaneously quantitating IgG responses to several Pn serotypes while conserving serum volumes and laboratory testing time.

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

10.1158/1538-7445.am2013-4541 ◽

2013 ◽

Author(s):

Carsten Jacobi ◽

Alain Schilb ◽

Yusuke Kawase ◽

Yasuyuki Kirii ◽

Brigitte Boldyreff

Keyword(s):

Meso Scale Discovery ◽

Array Platform ◽

Highly Sensitive ◽

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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