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Sci. Pharm. 2011, 79(3), 569-582; doi:10.3797/scipharm.1104-18

Validation of a Flow Cytometry Based Binding Assay for Evaluation of Monoclonal Antibody Recognizing EGF Receptor

Center of Molecular Immunology, 216 St and 15 Ave. Atabey, Playa. POBox 16040, Havana 11600, Cuba
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Received: 25 April 2011 / Accepted: 3 July 2011 / Published: 3 July 2011
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Abstract

An ideal test used to characterize a product must be appropriate for the measurement of product quality, manufacturing consistency, product stability, and comparability studies. Flow cytometry has been successfully applied to the examination of antibodies and receptors on membrane surfaces; however, to date, the analytical validation of cytometry based assays is limited. Here we report on the validation of a flow cytometry-based assay used in the evaluation of nimotuzumab binding to cells over-expressing EGFR on cell surface. The assay was validated by examining, assay robustness, specificity, repeatability and intermediate precision. The assay was highly specific, robust for all studied factors except for cell fixation with 1% paraformaldehyde and met criteria for precision with RSD < 2%. In addition the assay has stability-indicating proper-ties evidenced by the ability to detect changes in mAb degraded samples. Most importantly, the assay demonstrated to be useful for its intended use.
Keywords: Binding Assays; Method validation; Monoclonal antibody; Nimotuzumab Binding Assays; Method validation; Monoclonal antibody; Nimotuzumab
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

CEDEÑO-ARIAS, M.; SÁNCHEZ-RAMÍREZ, J.; BLANCO-SANTANA, R.; RENGIFO-CALZADO, E. Validation of a Flow Cytometry Based Binding Assay for Evaluation of Monoclonal Antibody Recognizing EGF Receptor. Sci. Pharm. 2011, 79, 569-582.

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