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bioRxiv · 10.1101/2024.10.31.621446

Development of an ELISA assay for EV quantification to assist the large-scale EV manufacture

Zhou, X.·Wang, W.·Zhang, L.·Yang, S.·Peng, X.·Zhang, X.·Zhao, J.·Xu, K.·He, X.
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Abstract

Accurate EV quantitation is essential for ensuring quality, consistency, and safety during large-scale extracellular vesicle (EV) manufacturing. In the upstream phase, EV quantitation allows for the monitoring of cell culture conditions that impact EV yield and quality, while in the downstream phase, it helps to control the efficiency and purity of EV purification. Nanoparticle Tracking Analysis is the most commonly used method for EV quantification, while it faces significant limitations due to interference from nanosize contaminants like protein aggregates, especially in crude samples (e.g. cell culture media). To address this issue, we developed a highly specific and accurate ELISA assay that quantifies EVs even in crude samples. With ultra-pure EV standard samples, this assay showed reliable quantitative result of EV detection to support method development as well as in-process control of large-scale EV manufacture. The detection range of this assay is from 4.1E7 to 3E10 EVs/mL, with an LOD of 1.04E7 EVs/mL and an LOQ of 3.21E7 EVs/mL. We therefore developed this assay into a testing kit and demonstrated that this EV quantification ELISA kit is capable of ensuring minimal interference from impurities and supporting the process development and in-process control in EV production.

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Zhou, X., Wang, W., Zhang, L., Yang, S., Peng, X., Zhang, X., Zhao, J., Xu, K., He, X.. 2024-11-03. Development of an ELISA assay for EV quantification to assist the large-scale EV manufacture. https://doi.org/10.1101/2024.10.31.621446

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