Table of Contents
International Scholarly Research Notices
Volume 2014, Article ID 487101, 7 pages
http://dx.doi.org/10.1155/2014/487101
Research Article

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

1Department of Quality Control, Center of Molecular Immunology, 11600 Havana, Cuba
2Biochemistry Laboratory, Quality Control Department, Center of Molecular Immunology, 11600 Havana, Cuba

Received 15 May 2014; Accepted 30 July 2014; Published 27 October 2014

Academic Editor: Qi Zhang

Copyright © 2014 Lesly Paradina Fernández et al. 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.

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.