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Published on: September 10, 2018
Development and validation of a cell-based assay to determine the bioactivity of antibodies against CD39 targets
Gangling Xu1, Yongbo Ni1, Chunbo Cui1
1National Institutes for Food and Drug Control, Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech, Beijing 102629, China; State Key Laboratory of Drug Regulatory Science, Beijing 102629, China.
Abstract:
CD39 is a novel therapeutic target involved in antitumor immunity that can be manipulated through the ATP-adenosine signaling pathway. As a promising cancer treatment option, several drugs targeting CD39 have recently progressed into clinical trials. In this study, we describe the development and validation of a novel method to evaluate the relative bioactivity of anti-CD39 antibodies based on CD39-expressing CHO-K1-huCD39-10B6 cells. The method uses a commercially available CellTiter-Glo luminescent cell viability assay kit to generate luciferase signals, thereby providing convenient and rapid sample analysis. The method was comprehensively validated following the ICH Q2 guidelines, and the results showed that this method demonstrated high specificity, accuracy, precision, and linearity, with a range of 60-140 %. Additionally, the CHO-K1-huCD39-10B6 cell line exhibited relative stability and robustness within 15 generations. Therefore this method has potential use in anti-CD39 antibody bioactivity assays for lot release and stability studies.
Insights
A new method was developed to assess anti-CD39 antibody bioactivity using CD39-expressing cells and a luminescence assay. This validated approach offers a reliable tool for cancer immunotherapy drug development and quality control.
Area of Science:
- Immunology
- Biotechnology
- Pharmacology
Background:
- CD39 is a key target in antitumor immunity, modulating the ATP-adenosine pathway.
- Anti-CD39 therapies are emerging as promising cancer treatment options, with several drugs in clinical trials.
Purpose of the Study:
- To develop and validate a novel method for evaluating the relative bioactivity of anti-CD39 antibodies.
- To establish a robust assay for quality control of anti-CD39 therapeutics.
Main Methods:
- Utilized CD39-expressing CHO-K1-huCD39-10B6 cells for antibody bioactivity assessment.
- Employed a commercially available CellTiter-Glo luminescent cell viability assay for signal generation.
- Validated the method according to ICH Q2 guidelines for specificity, accuracy, precision, and linearity.
Main Results:
- The developed method demonstrated high specificity, accuracy, precision, and linearity (60-140%).
- The CHO-K1-huCD39-10B6 cell line showed stability and robustness over 15 generations.
- The assay provides convenient and rapid sample analysis.
Conclusions:
- The novel bioactivity assay is suitable for anti-CD39 antibody characterization.
- This method can be applied for lot release and stability studies of anti-CD39 therapeutics.
- The validated assay supports the advancement of CD39-targeted cancer immunotherapies.
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