title: Development and Validation of an Affinity Chromatography-Protein G Method for IgG Quantification.
abstract: Nimotuzumab, an IgG that recognizes the epidermal growth factor receptor (EGF-R) overexpressed in some tumors, is used in the treatment of advanced head and neck cancer. For the quantification of this protein in cell culture supernatants, protein G-HPLC affinity chromatography is used due to its high affinity and specificity for antibodies of this class. The technique relies on the comparison of the area under the curve of the elution peak of the samples to be evaluated versus to a calibration curve of well-known concentrations and was validated by assessment of its robustness, specificity, repeatability, intermediate precision, accuracy, linearity, limit of detection, limit of quantification, and range. According to results of the study all validation parameters fulfilled the preestablished acceptance criteria and demonstrated the feasibility of the assay for the analysis of samples of cell culture supernatant as well as drug product.
authors.affiliation: Department of Quality Control, Center of Molecular Immunology, 11600 Havana, Cuba; Biochemistry Laboratory, Quality Control Department, Center of Molecular Immunology, 11600 Havana, Cuba.
authors.forename: Lesly
authors.initials: L
authors.lastname: Paradina Fernández
authors.affiliation: Department of Quality Control, Center of Molecular Immunology, 11600 Havana, Cuba.
authors.forename: Loany
authors.initials: L
authors.lastname: Calvo
authors.affiliation: Department of Quality Control, Center of Molecular Immunology, 11600 Havana, Cuba.
authors.forename: Lisel
authors.initials: L
authors.lastname: Viña