Validation of a flow cytometry‐based assay to assess C5aR receptor occupancy on neutrophils and monocytes for use in drug development

Quadrini, Karen J.; Hegelund, Anne Charlotte; Cortes, Kasia E.; Xue, Chengsen; Kennelly, Susan M.; Ji, Hong; Högerkorp, Carl-Magnus; Closkey, Thomas W. Mc

doi:10.1002/cyto.b.21260

Cited by 9 publications

(6 citation statements)

References 36 publications

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“…Standardization of the FC instrument is important for detecting RO [25][26][27]. Many aspects influence the measurement of RO of anti-PD-1 antibody drugs, including sample collection, storage and processing time, antibody reagent selection, sample processing standard operating procedures, instrument parameter setting and data analysis.…”

Section: Methods Validationmentioning

confidence: 99%

Receptor Occupancy Measurement of Anti-PD-1 Antibody Drugs in Support of Clinical Trials

Fang

Dong

et al. 2019

Bioanalysis

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Aim: The reliable measurement of receptor occupancy (RO) provides informative data for efficacy and safety evaluation. This study aimed to assess factors affecting RO measurement of anti-PD-1 antibodies in clinical studies. Materials & methods: RO performance was assessed using different T-cell activation markers measured by flow cytometry. The validated methodology was then used in support of a clinical study. Results: The optimized active cell population was comprised of CD45RO+ or CD45RA− T cells. The bioanalytical method was validated for inter- and intra-assay precision (coefficient of variation ≤30%) and sample storage stability for 3 days. Consistent RO saturation was observed in Phase Ia clinical trial, although receptor regulation appeared to be different. The formation of anti-drug antibodies had markedly influenced pharmacokinetics and RO. Conclusion: RO measurement in combination with pharmacokinetics and anti-drug antibodies data could allow the integrated evaluation and better understanding of efficacy and safety.

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Section: Methods Validationmentioning

confidence: 99%

Receptor Occupancy Measurement of Anti-PD-1 Antibody Drugs in Support of Clinical Trials

Fang

Dong

et al. 2019

Bioanalysis

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“…The flow cytometry assay was able to analyze a high number of cells and obtain an accurate and objective reading with a considerable reduction of the analysis time. This study expands the use of flow cytometry technology in drug development and toxicology .…”

Section: The Use Of Flow Cytometry In Infectious Disease and Toxicologymentioning

confidence: 90%

Issue Highlights—September 2018 (94B5)

Wang

2018

Cytometry Part B Clinical

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“…These measurements are particularly important given the prevalence of anti‐drug antibodies in xenogeneic species (Thway et al, 2013). When used as a pharmacodynamic (PD) measurement, ROA data may be combined with pharmacokinetic (PK) data for modeling in order to optimize clinical trial design, guide dose selection, and determine the duration and extent of target saturation (Fisher et al, 2016; Quadrini et al, 2016; Spilker, Singh, & Vicini, 2016; Sternebring et al, 2016; Wyant, Estevam, Yang, & Rosario, 2016). When integrated with anti‐drug‐antibody (ADA) results, ROA data are used to assess the impact of ADA on receptor engagement and to monitor the association of long‐term receptor binding with adverse events.…”

Section: Introductionmentioning

confidence: 99%

“…clinical trial design, guide dose selection, and determine the duration and extent of target saturation (Fisher et al, 2016;Quadrini et al, 2016;Spilker, Singh, & Vicini, 2016;Sternebring et al, 2016;Wyant, Estevam, Yang, & Rosario, 2016). When integrated with anti-drugantibody (ADA) results, ROA data are used to assess the impact of ADA on receptor engagement and to monitor the association of longterm receptor binding with adverse events.…”

mentioning

confidence: 99%

Best practices for optimization and validation of flow cytometry‐based receptor occupancy assays

Hilt

Sun

McCloskey³

et al. 2020

Cytometry Part B Clinical

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In the development of therapeutic compounds that bind cell surface molecules, it is critical to demonstrate the extent to which the drug engages its target. For cell‐associated targets, flow cytometry is well‐suited to monitor drug‐to‐target engagement through receptor occupancy assays (ROA). The technology allows for the identification of specific cell subsets within heterogeneous populations and the detection of nonabundant cellular antigens. There are numerous challenges in the design, development, and implementation of robust ROA. Among the most difficult challenges are situations where there is receptor modulation or when the target‐antigen is expressed at low levels. When the therapeutic molecules are bi‐specific and bind multiple targets, these challenges are increased. This manuscript discusses the challenges and proposes best practices for designing, optimizing, and validating ROA.

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Validation of a flow cytometry‐based assay to assess C5aR receptor occupancy on neutrophils and monocytes for use in drug development

Cited by 9 publications

References 36 publications

Receptor Occupancy Measurement of Anti-PD-1 Antibody Drugs in Support of Clinical Trials

Receptor Occupancy Measurement of Anti-PD-1 Antibody Drugs in Support of Clinical Trials

Issue Highlights—September 2018 (94B5)

Best practices for optimization and validation of flow cytometry‐based receptor occupancy assays

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