TY - JOUR
T1 - Batch-to-batch reproducibility of Transferon™
AU - Medina-Rivero, Emilio
AU - Merchand-Reyes, Giovanna
AU - Pavón, Lenin
AU - Vázquez-Leyva, Said
AU - Pérez-Sánchez, Gilberto
AU - Salinas-Jazmín, Nohemí
AU - Estrada-Parra, Sergio
AU - Velasco-Velázquez, Marco
AU - Pérez-Tapia, Sonia Mayra
PY - 2014/1/25
Y1 - 2014/1/25
N2 - Human dialyzable leukocyte extracts (DLEs) are heterogeneous mixtures of low-molecular-weight peptides that modulate immune responses in various diseases. Due their complexity, standardized methods to identify their physicochemical properties and determine that production batches are biologically active must be established. We aimed to develop and validate a size exclusion ultra performance chromatographic (SE-UPLC) method to characterize Transferon™, a DLE that is produced under good manufacturing practices (GMPs). We analyzed an internal human DLE standard and 10 representative batches of Transferon™, all of which had a chromatographic profile characterized by 8 main peaks and a molecular weight range between 17.0 and 0.2. kDa. There was high homogeneity between batches with regard to retention times and area percentages, varying by less than 0.2% and 30%, respectively, and the control chart was within 3 standard deviations. To analyze the biological activity of the batches, we studied the ability of Transferon™ to stimulate IFN-γ production in vitro. Transferon™ consistently induced IFN-γ production in Jurkat cells, demonstrating that this method can be included as a quality control step in releasing Transferon™ batches. Because all analyzed batches complied with the quality attributes that were evaluated, we conclude that the DLE Transferon™ is produced with high homogeneity.
AB - Human dialyzable leukocyte extracts (DLEs) are heterogeneous mixtures of low-molecular-weight peptides that modulate immune responses in various diseases. Due their complexity, standardized methods to identify their physicochemical properties and determine that production batches are biologically active must be established. We aimed to develop and validate a size exclusion ultra performance chromatographic (SE-UPLC) method to characterize Transferon™, a DLE that is produced under good manufacturing practices (GMPs). We analyzed an internal human DLE standard and 10 representative batches of Transferon™, all of which had a chromatographic profile characterized by 8 main peaks and a molecular weight range between 17.0 and 0.2. kDa. There was high homogeneity between batches with regard to retention times and area percentages, varying by less than 0.2% and 30%, respectively, and the control chart was within 3 standard deviations. To analyze the biological activity of the batches, we studied the ability of Transferon™ to stimulate IFN-γ production in vitro. Transferon™ consistently induced IFN-γ production in Jurkat cells, demonstrating that this method can be included as a quality control step in releasing Transferon™ batches. Because all analyzed batches complied with the quality attributes that were evaluated, we conclude that the DLE Transferon™ is produced with high homogeneity.
KW - Dialyzable leukocyte extracts
KW - IFN-gamma
KW - Quality specifications
KW - Transferon
KW - UPLC
UR - http://www.scopus.com/inward/record.url?scp=84885070260&partnerID=8YFLogxK
U2 - 10.1016/j.jpba.2013.09.004
DO - 10.1016/j.jpba.2013.09.004
M3 - Artículo
C2 - 24099727
AN - SCOPUS:84885070260
SN - 0731-7085
VL - 88
SP - 289
EP - 294
JO - Journal of Pharmaceutical and Biomedical Analysis
JF - Journal of Pharmaceutical and Biomedical Analysis
ER -