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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...

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Autores principales: Schmohl, Joerg U., Todhunter, Deborah, Oh, Seung, Vallera, Daniel A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2015
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.
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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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