Site-specific antigen-adjuvant conjugation using cell-free protein synthesis enhances antigen presentation and CD8+ T-cell response

Abstract 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...

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Autores principales:	Adam M. Weiss, Jainu Ajit, Tyler J. Albin, Neeraj Kapoor, Shilpa Maroju, Aym Berges, Lucy Pill, Jeff Fairman, Aaron P. Esser-Kahn
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/6dea9876b850448097ee9e13c5ae1009
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spelling	oai:doaj.org-article:6dea9876b850448097ee9e13c5ae10092021-12-02T13:17:42ZSite-specific antigen-adjuvant conjugation using cell-free protein synthesis enhances antigen presentation and CD8+ T-cell response10.1038/s41598-021-85709-12045-2322https://doaj.org/article/6dea9876b850448097ee9e13c5ae10092021-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-85709-1https://doaj.org/toc/2045-2322Abstract 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.Adam M. WeissJainu AjitTyler J. AlbinNeeraj KapoorShilpa MarojuAym BergesLucy PillJeff FairmanAaron P. Esser-KahnNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Adam M. Weiss Jainu Ajit Tyler J. Albin Neeraj Kapoor Shilpa Maroju Aym Berges Lucy Pill Jeff Fairman Aaron P. Esser-Kahn Site-specific antigen-adjuvant conjugation using cell-free protein synthesis enhances antigen presentation and CD8+ T-cell response
description	Abstract 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	article
author	Adam M. Weiss Jainu Ajit Tyler J. Albin Neeraj Kapoor Shilpa Maroju Aym Berges Lucy Pill Jeff Fairman Aaron P. Esser-Kahn
author_facet	Adam M. Weiss Jainu Ajit Tyler J. Albin Neeraj Kapoor Shilpa Maroju Aym Berges Lucy Pill Jeff Fairman Aaron P. Esser-Kahn
author_sort	Adam M. Weiss
title	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_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_sort	site-specific antigen-adjuvant conjugation using cell-free protein synthesis enhances antigen presentation and cd8+ t-cell response
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/6dea9876b850448097ee9e13c5ae1009
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Site-specific antigen-adjuvant conjugation using cell-free protein synthesis enhances antigen presentation and CD8+ T-cell response

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