Development of Functional Antibodies Directed to Human Dialyzable Leukocyte Extract (Transferon®)

Transferon® is an immunomodulator made of a complex mixture of peptides from human dialyzable leucocyte extracts (hDLEs). Development of surrogate antibodies directed to hDLE is an indispensable tool for studies during process control and preclinical trials. These antibodies are fundamental for diff...

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Main Authors: Gabriela Mellado-Sánchez, Juan José Lázaro-Rodríguez, Sandra Avila, Luis Vallejo-Castillo, Said Vázquez-Leyva, Gregorio Carballo-Uicab, Marco Velasco-Velázquez, Emilio Medina-Rivero, Lenin Pavón, Rommel Chacón-Salinas, Sonia Mayra Pérez-Tapia
Format: Article
Language:English
Published: Wiley 2019-01-01
Series:Journal of Immunology Research
Online Access:http://dx.doi.org/10.1155/2019/2754920
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author Gabriela Mellado-Sánchez
Juan José Lázaro-Rodríguez
Sandra Avila
Luis Vallejo-Castillo
Said Vázquez-Leyva
Gregorio Carballo-Uicab
Marco Velasco-Velázquez
Emilio Medina-Rivero
Lenin Pavón
Rommel Chacón-Salinas
Sonia Mayra Pérez-Tapia
author_facet Gabriela Mellado-Sánchez
Juan José Lázaro-Rodríguez
Sandra Avila
Luis Vallejo-Castillo
Said Vázquez-Leyva
Gregorio Carballo-Uicab
Marco Velasco-Velázquez
Emilio Medina-Rivero
Lenin Pavón
Rommel Chacón-Salinas
Sonia Mayra Pérez-Tapia
author_sort Gabriela Mellado-Sánchez
collection DOAJ
description Transferon® is an immunomodulator made of a complex mixture of peptides from human dialyzable leucocyte extracts (hDLEs). Development of surrogate antibodies directed to hDLE is an indispensable tool for studies during process control and preclinical trials. These antibodies are fundamental for different analytical approaches, such as identity test and drug quantitation, as well as to characterize its pharmacokinetic and mechanisms of action. A previous murine study showed the inability of the peptides of Transferon® to induce antibody production by themselves; therefore, in this work, two approaches were tested to increase its immunogenicity: chemical conjugation of the peptides of Transferon® to carrier proteins and the use of a rabbit model. Bioconjugates were generated with Keyhole Limpet Hemocyanin (KLH) or Bovine Serum Albumin (BSA) through maleimide-activated carrier proteins. BALB/c mice and New Zealand rabbits were immunized with Transferon® conjugated to KLH or nonconjugated Transferon®. Animals that were immunized with conjugated Transferon® showed significant production of antibodies as evinced by the recognition of Transferon®-BSA conjugate in ELISA assays. Moreover, rabbits showed higher antibody titers when compared with mice. Neither mouse nor rabbits developed antibodies when immunized with nonconjugated Transferon®. Interestingly, rabbit antibodies were able to partially block IL-2 production in Jurkat cells after costimulation with Transferon®. In conclusion, it is feasible to elicit specific and functional antibodies anti-hDLE with different potential uses during the life cycle of the product.
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spelling doaj-art-b39ff32291fc48699d74176ea6d223c92025-08-20T03:20:51ZengWileyJournal of Immunology Research2314-88612314-71562019-01-01201910.1155/2019/27549202754920Development of Functional Antibodies Directed to Human Dialyzable Leukocyte Extract (Transferon®)Gabriela Mellado-Sánchez0Juan José Lázaro-Rodríguez1Sandra Avila2Luis Vallejo-Castillo3Said Vázquez-Leyva4Gregorio Carballo-Uicab5Marco Velasco-Velázquez6Emilio Medina-Rivero7Lenin Pavón8Rommel Chacón-Salinas9Sonia Mayra Pérez-Tapia10Unidad de Desarrollo e Investigación en Bioprocesos (UDIBI), Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, CDMX, MexicoUnidad de Desarrollo e Investigación en Bioprocesos (UDIBI), Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, CDMX, MexicoUnidad de Desarrollo e Investigación en Bioprocesos (UDIBI), Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, CDMX, MexicoUnidad de Desarrollo e Investigación en Bioprocesos (UDIBI), Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, CDMX, MexicoUnidad de Desarrollo e Investigación en Bioprocesos (UDIBI), Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, CDMX, MexicoUnidad de Desarrollo e Investigación en Bioprocesos (UDIBI), Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, CDMX, MexicoDepartamento de Farmacología y Unidad Periférica de Investigación en Biomedicina Translacional (CMN 20 de noviembre, ISSSTE), Facultad de Medicina, Universidad Nacional Autónoma de México, Ciudad Universitaria, CDMX, MexicoUnidad de Desarrollo e Investigación en Bioprocesos (UDIBI), Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, CDMX, MexicoLaboratorio de Psicoinmunología, Instituto Nacional de Psiquiatría Ramón de la Fuente, CDMX, MexicoUnidad de Desarrollo e Investigación en Bioprocesos (UDIBI), Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, CDMX, MexicoUnidad de Desarrollo e Investigación en Bioprocesos (UDIBI), Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, CDMX, MexicoTransferon® is an immunomodulator made of a complex mixture of peptides from human dialyzable leucocyte extracts (hDLEs). Development of surrogate antibodies directed to hDLE is an indispensable tool for studies during process control and preclinical trials. These antibodies are fundamental for different analytical approaches, such as identity test and drug quantitation, as well as to characterize its pharmacokinetic and mechanisms of action. A previous murine study showed the inability of the peptides of Transferon® to induce antibody production by themselves; therefore, in this work, two approaches were tested to increase its immunogenicity: chemical conjugation of the peptides of Transferon® to carrier proteins and the use of a rabbit model. Bioconjugates were generated with Keyhole Limpet Hemocyanin (KLH) or Bovine Serum Albumin (BSA) through maleimide-activated carrier proteins. BALB/c mice and New Zealand rabbits were immunized with Transferon® conjugated to KLH or nonconjugated Transferon®. Animals that were immunized with conjugated Transferon® showed significant production of antibodies as evinced by the recognition of Transferon®-BSA conjugate in ELISA assays. Moreover, rabbits showed higher antibody titers when compared with mice. Neither mouse nor rabbits developed antibodies when immunized with nonconjugated Transferon®. Interestingly, rabbit antibodies were able to partially block IL-2 production in Jurkat cells after costimulation with Transferon®. In conclusion, it is feasible to elicit specific and functional antibodies anti-hDLE with different potential uses during the life cycle of the product.http://dx.doi.org/10.1155/2019/2754920
spellingShingle Gabriela Mellado-Sánchez
Juan José Lázaro-Rodríguez
Sandra Avila
Luis Vallejo-Castillo
Said Vázquez-Leyva
Gregorio Carballo-Uicab
Marco Velasco-Velázquez
Emilio Medina-Rivero
Lenin Pavón
Rommel Chacón-Salinas
Sonia Mayra Pérez-Tapia
Development of Functional Antibodies Directed to Human Dialyzable Leukocyte Extract (Transferon®)
Journal of Immunology Research
title Development of Functional Antibodies Directed to Human Dialyzable Leukocyte Extract (Transferon®)
title_full Development of Functional Antibodies Directed to Human Dialyzable Leukocyte Extract (Transferon®)
title_fullStr Development of Functional Antibodies Directed to Human Dialyzable Leukocyte Extract (Transferon®)
title_full_unstemmed Development of Functional Antibodies Directed to Human Dialyzable Leukocyte Extract (Transferon®)
title_short Development of Functional Antibodies Directed to Human Dialyzable Leukocyte Extract (Transferon®)
title_sort development of functional antibodies directed to human dialyzable leukocyte extract transferon r
url http://dx.doi.org/10.1155/2019/2754920
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