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The Thai Journal of Pharmaceutical Sciences

Abstract

Nimotuzumab has been used for treating squamous cell carcinoma of the head and neck. Its efficacy is attributed to its mechanism of action, which involves the binding to the epidermal growth factor receptor. Ensuring the quality of nimotuzumab is crucial for its safety and efficacy. Utilizing a binding assay to measure its efficacy is a valid approach since it directly correlates with its potency. Here, we developed and validated the cell-based binding assay to determine the potency of nimotuzumab. A-431 cells and fluorescein isothiocyanate mouse anti-human immunoglobulin G antibody were used and the fluorescent intensity was measured to evaluate the binding activity. The binding assay conditions were optimized during the method development and the final method was fully validated. The binding method was established by optimizing the suitability and appropriate amount of the antibody, incubation time for binding assay, and the condition for cell washing. The results of specificity and the linearity range between 1.5 and 4.5 μg/ml of nimotuzumab were in the acceptable criteria. The recoveries of nimotuzumab at 75, 100, and 125% of the target concentration demonstrated that the method was accurate. The relative standard deviations for repeatability and intermediate precision were within the limit. The data revealed that the developed method was successfully validated. The developed and validated cellbased potency assay can be employed for the potency determination of nimotuzumab products marketed in Thailand and the method can later be adapted to evaluate biosimilar nimotuzumab that will be developed and registered in the foreseeable future.

DOI

10.56808/3027-7922.2910

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