Validation of a cell proliferation assay to assess the potency of a dialyzable leukocyte extract intended for batch release

Gregorio Carballo-Uicab, José E. Linares-Trejo, Gabriela Mellado-Sánchez, Carlos A. López-Morales, Marco Velasco-Velázquez, Lenin Pavón, Sergio Estrada-Parra, Sonia Mayra Pérez-Tapia, Emilio Medina-Rivero

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

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®.

Original languageEnglish
Article number3426
JournalMolecules
Volume24
Issue number19
DOIs
StatePublished - 20 Sep 2019

Keywords

  • Biological potency
  • Complex mixture of peptides
  • Development and validation
  • Dialyzable leukocyte extract
  • Quality specifications
  • Transferon®

Fingerprint

Dive into the research topics of 'Validation of a cell proliferation assay to assess the potency of a dialyzable leukocyte extract intended for batch release'. Together they form a unique fingerprint.

Cite this