TY - JOUR
T1 - Validation of a cell proliferation assay to assess the potency of a dialyzable leukocyte extract intended for batch release
AU - Carballo-Uicab, Gregorio
AU - Linares-Trejo, José E.
AU - Mellado-Sánchez, Gabriela
AU - López-Morales, Carlos A.
AU - Velasco-Velázquez, Marco
AU - Pavón, Lenin
AU - Estrada-Parra, Sergio
AU - Pérez-Tapia, Sonia Mayra
AU - Medina-Rivero, Emilio
N1 - Publisher Copyright:
© 2019 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2019/9/20
Y1 - 2019/9/20
N2 - Transferon® is a blood product with immunomodulatory properties constituted by a complex mixture of peptides obtained from a human dialyzable leukocyte extract (DLE). Due to its complex nature, it is necessary to demonstrate batch consistency in its biological activity. Potency is the quantitative measure of biological activity and is also a quality attribute of drugs. Here we developed and validated a proliferation assay using Jurkat cells exposed to azathioprine, which is intended to determine the potency of Transferon® according to international guidelines for pharmaceuticals. The assay showed a linear response (2.5 to 40 μg/mL), coefficients of variation from 0.7 to 13.6% demonstrated that the method is precise, while r2 = 0.97 between the nominal and measured values obtained from dilutional linearity showed that the method is accurate. We also demonstrated that the cell proliferation response was specific for Transferon® and was not induced by its vehicle nor by other peptide complex mixtures (glatiramer acetate or hydrolyzed collagen). The bioassay validated here was used to assess the relative potency of eight released batches of Transferon® with respect to a reference standard, showing consistent results. The collective information from the validation and the assessment of several batches indicate that the bioassay is suitable for the release of Transferon®.
AB - Transferon® is a blood product with immunomodulatory properties constituted by a complex mixture of peptides obtained from a human dialyzable leukocyte extract (DLE). Due to its complex nature, it is necessary to demonstrate batch consistency in its biological activity. Potency is the quantitative measure of biological activity and is also a quality attribute of drugs. Here we developed and validated a proliferation assay using Jurkat cells exposed to azathioprine, which is intended to determine the potency of Transferon® according to international guidelines for pharmaceuticals. The assay showed a linear response (2.5 to 40 μg/mL), coefficients of variation from 0.7 to 13.6% demonstrated that the method is precise, while r2 = 0.97 between the nominal and measured values obtained from dilutional linearity showed that the method is accurate. We also demonstrated that the cell proliferation response was specific for Transferon® and was not induced by its vehicle nor by other peptide complex mixtures (glatiramer acetate or hydrolyzed collagen). The bioassay validated here was used to assess the relative potency of eight released batches of Transferon® with respect to a reference standard, showing consistent results. The collective information from the validation and the assessment of several batches indicate that the bioassay is suitable for the release of Transferon®.
KW - Biological potency
KW - Complex mixture of peptides
KW - Development and validation
KW - Dialyzable leukocyte extract
KW - Quality specifications
KW - Transferon®
UR - http://www.scopus.com/inward/record.url?scp=85072552906&partnerID=8YFLogxK
U2 - 10.3390/molecules24193426
DO - 10.3390/molecules24193426
M3 - Artículo
C2 - 31547184
AN - SCOPUS:85072552906
SN - 1420-3049
VL - 24
JO - Molecules
JF - Molecules
IS - 19
M1 - 3426
ER -