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Validation of a Cell Proliferation Assay to Assess the Potency of a Dialyzable Leukocyte Extract Intended for Batch Release

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Unidad de Desarrollo e Investigación en Bioprocesos (UDIBI), Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Ciudad de Mexico 11340, Mexico
2
Departamento de Farmacología y Unidad Periférica de Investigación en Biomedicina Traslacional (CMN 20 de noviembre, ISSSTE), Facultad de Medicina, Universidad Nacional Autónoma de México, Ciudad Universitaria, Ciudad de Mexico 04510, Mexico
3
Laboratorio de Psicoinmunología, Dirección de Investigaciones en Neurociencias del Instituto Nacional de Psiquiatría Ramón de la Fuente, Cuida de Mexico 14370, Mexico
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Departamento de Inmunología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Ciudad de Mexico 11340, Mexico
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Laboratorio Nacional para Servicios Especializados de Investigación, Desarrollo e Innovación (I+D+i) para Farmacoquímicos y Biotecnológicos, LANSEIDI-FarBiotec-CONACyT, Ciudad de Mexico 11340, Mexico
*
Authors to whom correspondence should be addressed.
Academic Editors: In-Soo Yoon and Hyun-Jong Cho
Molecules 2019, 24(19), 3426; https://doi.org/10.3390/molecules24193426
Received: 28 August 2019 / Revised: 6 September 2019 / Accepted: 8 September 2019 / Published: 20 September 2019
(This article belongs to the Special Issue Method Development and Validation in Food and Pharmaceutical Analysis)
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®. View Full-Text
Keywords: dialyzable leukocyte extract; Transferon®; complex mixture of peptides; quality specifications; biological potency; development and validation dialyzable leukocyte extract; Transferon®; complex mixture of peptides; quality specifications; biological potency; development and validation
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MDPI and ACS Style

Carballo-Uicab, G.; Linares-Trejo, J.E.; Mellado-Sánchez, G.; López-Morales, C.A.; Velasco-Velázquez, M.; Pavón, L.; Estrada-Parra, S.; Pérez-Tapia, S.M.; Medina-Rivero, E. Validation of a Cell Proliferation Assay to Assess the Potency of a Dialyzable Leukocyte Extract Intended for Batch Release. Molecules 2019, 24, 3426.

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