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Site-specific antigen-adjuvant conjugation using cell-free protein synthesis enhances antigen presentation and CD8(+) T-cell response
Antigen-adjuvant conjugation is known to enhance antigen-specific T-cell production in vaccine models, but scalable methods are required to generate site-specific conjugation for clinical translation of this technique. We report the use of the cell-free protein synthesis (CFPS) platform as a rapid m...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Nature Publishing Group UK
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7973483/ https://www.ncbi.nlm.nih.gov/pubmed/33737644 http://dx.doi.org/10.1038/s41598-021-85709-1 |
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author | Weiss, Adam M. Ajit, Jainu Albin, Tyler J. Kapoor, Neeraj Maroju, Shilpa Berges, Aym Pill, Lucy Fairman, Jeff Esser-Kahn, Aaron P. |
author_facet | Weiss, Adam M. Ajit, Jainu Albin, Tyler J. Kapoor, Neeraj Maroju, Shilpa Berges, Aym Pill, Lucy Fairman, Jeff Esser-Kahn, Aaron P. |
author_sort | Weiss, Adam M. |
collection | PubMed |
description | Antigen-adjuvant conjugation is known to enhance antigen-specific T-cell production in vaccine models, but scalable methods are required to generate site-specific conjugation for clinical translation of this technique. We report the use of the cell-free protein synthesis (CFPS) platform as a rapid method to produce large quantities (> 100 mg/L) of a model antigen, ovalbumin (OVA), with site-specific incorporation of p-azidomethyl-l-phenylalanine (pAMF) at two solvent-exposed sites away from immunodominant epitopes. Using copper-free click chemistry, we conjugated CpG oligodeoxynucleotide toll-like receptor 9 (TLR9) agonists to the pAMF sites on the mutant OVA protein. The OVA-CpG conjugates demonstrate enhanced antigen presentation in vitro and increased antigen-specific CD8(+) T-cell production in vivo. Moreover, OVA-CpG conjugation reduced the dose of CpG needed to invoke antigen-specific T-cell production tenfold. These results highlight how site-specific conjugation and CFPS technology can be implemented to produce large quantities of covalently-linked antigen-adjuvant conjugates for use in clinical vaccines. |
format | Online Article Text |
id | pubmed-7973483 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-79734832021-03-19 Site-specific antigen-adjuvant conjugation using cell-free protein synthesis enhances antigen presentation and CD8(+) T-cell response Weiss, Adam M. Ajit, Jainu Albin, Tyler J. Kapoor, Neeraj Maroju, Shilpa Berges, Aym Pill, Lucy Fairman, Jeff Esser-Kahn, Aaron P. Sci Rep Article Antigen-adjuvant conjugation is known to enhance antigen-specific T-cell production in vaccine models, but scalable methods are required to generate site-specific conjugation for clinical translation of this technique. We report the use of the cell-free protein synthesis (CFPS) platform as a rapid method to produce large quantities (> 100 mg/L) of a model antigen, ovalbumin (OVA), with site-specific incorporation of p-azidomethyl-l-phenylalanine (pAMF) at two solvent-exposed sites away from immunodominant epitopes. Using copper-free click chemistry, we conjugated CpG oligodeoxynucleotide toll-like receptor 9 (TLR9) agonists to the pAMF sites on the mutant OVA protein. The OVA-CpG conjugates demonstrate enhanced antigen presentation in vitro and increased antigen-specific CD8(+) T-cell production in vivo. Moreover, OVA-CpG conjugation reduced the dose of CpG needed to invoke antigen-specific T-cell production tenfold. These results highlight how site-specific conjugation and CFPS technology can be implemented to produce large quantities of covalently-linked antigen-adjuvant conjugates for use in clinical vaccines. Nature Publishing Group UK 2021-03-18 /pmc/articles/PMC7973483/ /pubmed/33737644 http://dx.doi.org/10.1038/s41598-021-85709-1 Text en © The Author(s) 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Weiss, Adam M. Ajit, Jainu Albin, Tyler J. Kapoor, Neeraj Maroju, Shilpa Berges, Aym Pill, Lucy Fairman, Jeff Esser-Kahn, Aaron P. Site-specific antigen-adjuvant conjugation using cell-free protein synthesis enhances antigen presentation and CD8(+) T-cell response |
title | Site-specific antigen-adjuvant conjugation using cell-free protein synthesis enhances antigen presentation and CD8(+) T-cell response |
title_full | Site-specific antigen-adjuvant conjugation using cell-free protein synthesis enhances antigen presentation and CD8(+) T-cell response |
title_fullStr | Site-specific antigen-adjuvant conjugation using cell-free protein synthesis enhances antigen presentation and CD8(+) T-cell response |
title_full_unstemmed | Site-specific antigen-adjuvant conjugation using cell-free protein synthesis enhances antigen presentation and CD8(+) T-cell response |
title_short | Site-specific antigen-adjuvant conjugation using cell-free protein synthesis enhances antigen presentation and CD8(+) T-cell response |
title_sort | site-specific antigen-adjuvant conjugation using cell-free protein synthesis enhances antigen presentation and cd8(+) t-cell response |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7973483/ https://www.ncbi.nlm.nih.gov/pubmed/33737644 http://dx.doi.org/10.1038/s41598-021-85709-1 |
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