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Mutagenic Deimmunization of Diphtheria Toxin for Use in Biologic Drug Development
Background: Targeted toxins require multiple treatments and therefore must be deimmunized. We report a method of protein deimmunization based on the point mutation of highly hydrophilic R, K, D, E, and Q amino acids on the molecular surface of truncated diphtheria-toxin (DT390). Methods: Based on th...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4626721/ https://www.ncbi.nlm.nih.gov/pubmed/26473923 http://dx.doi.org/10.3390/toxins7104067 |
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author | Schmohl, Joerg U. Todhunter, Deborah Oh, Seung Vallera, Daniel A. |
author_facet | Schmohl, Joerg U. Todhunter, Deborah Oh, Seung Vallera, Daniel A. |
author_sort | Schmohl, Joerg U. |
collection | PubMed |
description | Background: Targeted toxins require multiple treatments and therefore must be deimmunized. We report a method of protein deimmunization based on the point mutation of highly hydrophilic R, K, D, E, and Q amino acids on the molecular surface of truncated diphtheria-toxin (DT390). Methods: Based on their surface position derived from an X-ray-crystallographic model, residues were chosen for point mutation that were located in prominent positions on the molecular surface and away from the catalytic site. Mice were immunized with a targeted toxin containing either a mutated DT390 containing seven critical point mutations or the non-mutated parental toxin form. Results: Serum analysis revealed a significant 90% reduction in anti-toxin antibodies in mice immunized with the mutant, but not the parental drug form despite multiple immunizations. The experiment was repeated in a second strain of mice with a different MHC-haplotype to address whether point mutation removed T or B cell epitopes. Findings were identical indicating that B cell epitopes were eliminated from DT. The mutant drug form lost only minimal activity in vitro as well as in vivo. Conclusion: These findings indicate that this method may be effective for deimmunizing of other proteins and that discovery of a deimmunized form of DT may lead to the development of more effective targeted toxin. |
format | Online Article Text |
id | pubmed-4626721 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-46267212015-11-12 Mutagenic Deimmunization of Diphtheria Toxin for Use in Biologic Drug Development Schmohl, Joerg U. Todhunter, Deborah Oh, Seung Vallera, Daniel A. Toxins (Basel) Article Background: Targeted toxins require multiple treatments and therefore must be deimmunized. We report a method of protein deimmunization based on the point mutation of highly hydrophilic R, K, D, E, and Q amino acids on the molecular surface of truncated diphtheria-toxin (DT390). Methods: Based on their surface position derived from an X-ray-crystallographic model, residues were chosen for point mutation that were located in prominent positions on the molecular surface and away from the catalytic site. Mice were immunized with a targeted toxin containing either a mutated DT390 containing seven critical point mutations or the non-mutated parental toxin form. Results: Serum analysis revealed a significant 90% reduction in anti-toxin antibodies in mice immunized with the mutant, but not the parental drug form despite multiple immunizations. The experiment was repeated in a second strain of mice with a different MHC-haplotype to address whether point mutation removed T or B cell epitopes. Findings were identical indicating that B cell epitopes were eliminated from DT. The mutant drug form lost only minimal activity in vitro as well as in vivo. Conclusion: These findings indicate that this method may be effective for deimmunizing of other proteins and that discovery of a deimmunized form of DT may lead to the development of more effective targeted toxin. MDPI 2015-10-10 /pmc/articles/PMC4626721/ /pubmed/26473923 http://dx.doi.org/10.3390/toxins7104067 Text en © 2015 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Schmohl, Joerg U. Todhunter, Deborah Oh, Seung Vallera, Daniel A. Mutagenic Deimmunization of Diphtheria Toxin for Use in Biologic Drug Development |
title | Mutagenic Deimmunization of Diphtheria Toxin for Use in Biologic Drug Development |
title_full | Mutagenic Deimmunization of Diphtheria Toxin for Use in Biologic Drug Development |
title_fullStr | Mutagenic Deimmunization of Diphtheria Toxin for Use in Biologic Drug Development |
title_full_unstemmed | Mutagenic Deimmunization of Diphtheria Toxin for Use in Biologic Drug Development |
title_short | Mutagenic Deimmunization of Diphtheria Toxin for Use in Biologic Drug Development |
title_sort | mutagenic deimmunization of diphtheria toxin for use in biologic drug development |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4626721/ https://www.ncbi.nlm.nih.gov/pubmed/26473923 http://dx.doi.org/10.3390/toxins7104067 |
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