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

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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Comparison of competitive ligand-binding assay and bioassay formats for the measurement of neutralizing antibodies to protein therapeutics

Journal of Pharmaceutical and Biomedical Analysis ◽

10.1016/j.jpba.2010.08.029 ◽

2011 ◽

Vol 54 (2) ◽

pp. 351-358 ◽

Cited By ~ 29

Author(s):

Deborah Finco ◽

Daniel Baltrukonis ◽

Adrienne Clements-Egan ◽

Kathy Delaria ◽

George R. Gunn ◽

...

Keyword(s):

Ligand Binding ◽

Neutralizing Antibodies ◽

Binding Assay ◽

Protein Therapeutics ◽

Ligand Binding Assay ◽

Competitive Ligand

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A competitive ligand-binding assay for the detection of neutralizing antibodies against dostarlimab (TSR-042)

AAPS Open ◽

10.1186/s41120-021-00039-w ◽

2021 ◽

Vol 7 (1) ◽

Author(s):

Xiaolong Tom Zhang ◽

Hong Chen ◽

Weiping Shao ◽

Zhongping John Lin ◽

Murad Melhem ◽

...

Keyword(s):

Ligand Binding ◽

Human Serum ◽

Neutralizing Antibodies ◽

Binding Assay ◽

False Positive Rate ◽

Ligand Binding Assay ◽

Drug Interference ◽

Hook Effect ◽

System Suitability ◽

Competitive Ligand

AbstractDostarlimab is a humanized anti–PD-1 monoclonal antibody. Dostarlimab (JEMPERLI; TSR-042) was recently approved in the USA and in the EU. The presence of neutralizing antibodies (NAbs) is a cause for concern because they block the therapeutic function of the antibody and reduce drug efficacy. Therefore, programs developing therapeutic biologics need to develop and validate assays that adequately assess the presence of NAbs in the serum of patients treated with biologic therapies. Presented here is the development and validation of a competitive ligand-binding assay that specifically detects anti-dostarlimab NAbs in human serum. Precision, sensitivity, hook effect, selectivity, assay robustness, stabilities, and system suitability were evaluated. In addition, drug tolerance of the assay with the implementation of a drug removal process was investigated. The cut point factor for the detection of NAbs in human serum at a 1% false-positive rate was determined. The assay’s precision, sensitivity, hook effect, selectivity, robustness, and drug interference were tested and found to be acceptable. With system suitability and stability established, this assay has been used to evaluate NAbs to guide the development of dostarlimab.Trial registration: Clinicaltrials.gov, NCT02715284. Registered 9 March 2016

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Development of an Meso Scale Discovery ligand-binding assay for measurement of free (drug-unbound) target in nonhuman primate serum

Bioanalysis ◽

10.4155/bio-2020-0307 ◽

2021 ◽

Author(s):

Yun Liu ◽

Ronald Robinson ◽

Thao Ung ◽

Chrysanthe Spais ◽

Justin Schreiber ◽

...

Keyword(s):

Ligand Binding ◽

Nonhuman Primate ◽

Binding Assay ◽

Free Form ◽

Ligand Binding Assay ◽

Target Engagement ◽

Meso Scale Discovery ◽

Drug Interference ◽

Assay Performance ◽

Meso Scale

Aim: To quantify the free form of a protein as a target-engagement biomarker in nonhuman primate serum, a Meso Scale Discovery ligand-binding assay was developed and qualified. Results: The initial assay produced an unexpected artifact when used to measure the free target in study samples dosed with drug. By using incurred study samples dosed with high drug levels to test assay performance, we developed an alternative assay that does not suffer from drug interference. Conclusion: Our work demonstrated that an assay designed to measure free target may not necessarily deliver reliable quantitation. In our case, incurred study samples dosed with drug proved to be useful in developing an alternative free assay that does not suffer from drug interference.

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