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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| Format: | Article |
| Language: | English |
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Wiley
2019-01-01
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| 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. |
| format | Article |
| id | doaj-art-b39ff32291fc48699d74176ea6d223c9 |
| institution | DOAJ |
| issn | 2314-8861 2314-7156 |
| language | English |
| publishDate | 2019-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Immunology Research |
| 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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