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Development of a VHH-Based Erythropoietin Quantification Assay.

Mol. Biotechnol.. 2015; 
KolStefan,KallehaugeThomas Beuchert,AdemaSimon,Herman
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Molecular Biology Reagents … The anti-EPO IgG was biotinylated using the EZ-Link biotinylation kit (Thermo Scientific, Rockford, IL, USA) with a ratio of biotin:antibody of 1 EPO dilution series were prepared using commercially bought EPO (Genscript, Piscataway, NJ, USA) and our in-house purified EPO at … Get A Quote

摘要

Erythropoietin (EPO) quantification during cell line selection and bioreactor cultivation has traditionally been performed with ELISA or HPLC. As these techniques suffer from several drawbacks, we developed a novel EPO quantification assay. A camelid single-domain antibody fragment directed against human EPO was evaluated as a capturing antibody in a label-free biolayer interferometry-based quantification assay. Human recombinant EPO can be specifically detected in Chinese hamster ovary cell supernatants in a sensitive and pH-dependent manner. This method enables rapid and robust quantification of EPO in a high-throughput setting.

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