Development and Validation of an Affinity Chromatography-Protein G Method for IgG Quantification.

Area of Science:

• Biochemistry
• Analytical Chemistry
• Pharmacology

Background:

• Nimotuzumab is an IgG antibody targeting the epidermal growth factor receptor (EGF-R).
• EGF-R is overexpressed in certain tumors, making nimotuzumab a therapeutic option for advanced head and neck cancer.
• Accurate quantification of nimotuzumab in biological samples is crucial for therapeutic monitoring and drug development.

Purpose of the Study:

• To develop and validate a robust Protein G-HPLC affinity chromatography method.
• To quantify nimotuzumab in cell culture supernatants and drug products.
• To ensure the reliability and feasibility of the assay for quality control and research.

Main Methods:

• Protein G-HPLC affinity chromatography was employed for antibody quantification.
• Method validation included assessment of robustness, specificity, repeatability, intermediate precision, accuracy, linearity, limit of detection, limit of quantification, and range.
• Quantification was based on comparing elution peak areas against a calibration curve of known concentrations.

Main Results:

• All pre-established validation parameters met the acceptance criteria.
• The method demonstrated high specificity and affinity for nimotuzumab.
• The assay proved feasible and reliable for analyzing nimotuzumab in cell culture supernatants and drug product.

Conclusions:

• The Protein G-HPLC method is a validated and reliable technique for nimotuzumab quantification.
• This assay is suitable for quality control and analysis of nimotuzumab in various pharmaceutical development stages.
• The validated method supports the consistent production and therapeutic use of nimotuzumab.