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

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  • Additional Information
    • Source:
      Publisher: Nature Publishing Group Country of Publication: England NLM ID: 101563288 Publication Model: Electronic Cited Medium: Internet ISSN: 2045-2322 (Electronic) Linking ISSN: 20452322 NLM ISO Abbreviation: Sci Rep Subsets: MEDLINE
    • Publication Information:
      Original Publication: London : Nature Publishing Group, copyright 2011-
    • Subject Terms:
      Antigen Presentation*; Adjuvants, Immunologic/*metabolism ; Antigens/*immunology ; CD8-Positive T-Lymphocytes/*immunology ; Mutant Proteins/*immunology ; Oligodeoxyribonucleotides/*immunology ; Ovalbumin/*immunology; Animals ; Antigen-Presenting Cells/immunology ; Antigens/genetics ; Cell-Free System ; Click Chemistry/methods ; HEK293 Cells ; Humans ; Mice ; Mice, Inbred C57BL ; Models, Animal ; Oligodeoxyribonucleotides/metabolism ; Oligodeoxyribonucleotides/pharmacology ; Ovalbumin/genetics ; Toll-Like Receptor 9/agonists ; Toll-Like Receptor 9/genetics ; Toll-Like Receptor 9/metabolism ; Transfection ; Vaccination/methods ; Vaccines, Conjugate/administration & dosage ; Vaccines, Conjugate/immunology ; Vaccines, Subunit/administration & dosage ; Vaccines, Subunit/immunology
    • 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.
    • References:
      Nat Biotechnol. 2010 Jun;28(6):573-9. (PMID: 20531338)
      PLoS One. 2017 Mar 29;12(3):e0173778. (PMID: 28355240)
      ChemMedChem. 2013 Mar;8(3):360-76. (PMID: 23316023)
      Proc Natl Acad Sci U S A. 2005 Oct 18;102(42):15190-4. (PMID: 16219698)
      Biochemistry. 2018 Feb 6;57(5):516-519. (PMID: 29323879)
      J Immunol. 2005 Apr 1;174(7):4373-80. (PMID: 15778402)
      Bioconjug Chem. 2019 Mar 20;30(3):561-567. (PMID: 30768262)
      Cancer Res. 2008 Jul 1;68(13):5390-6. (PMID: 18593941)
      Nature. 2005 Feb 24;433(7028):887-92. (PMID: 15711573)
      Vaccine. 2018 Jan 29;36(5):668-674. (PMID: 29289383)
      Cell Immunol. 2012;272(2):275-82. (PMID: 22051048)
      Science. 1996 Apr 5;272(5258):50-3. (PMID: 8600536)
      J Am Chem Soc. 2013 Feb 13;135(6):2044-7. (PMID: 23331082)
      Curr Opin Immunol. 2019 Aug;59:42-48. (PMID: 31003070)
      Vaccine. 2016 Dec 20;34(52):6672-6680. (PMID: 27836435)
      Trends Biotechnol. 2019 Apr;37(4):373-388. (PMID: 30470547)
      Biotechnol Adv. 2012 Sep-Oct;30(5):1185-94. (PMID: 22008973)
      Biotechnol Bioeng. 2011 Jul;108(7):1570-8. (PMID: 21337337)
      Science. 2001 Apr 20;292(5516):498-500. (PMID: 11313494)
      Bioconjug Chem. 2020 Nov 18;31(11):2585-2595. (PMID: 33151667)
      Nat Biotechnol. 2000 May;18(5):509-14. (PMID: 10802617)
      Eur J Immunol. 2002 Aug;32(8):2356-64. (PMID: 12209649)
    • Grant Information:
      T32 GM008720 United States GM NIGMS NIH HHS
    • Accession Number:
      0 (Adjuvants, Immunologic)
      0 (Antigens)
      0 (CPG-oligonucleotide)
      0 (Mutant Proteins)
      0 (Oligodeoxyribonucleotides)
      0 (Tlr9 protein, mouse)
      0 (Toll-Like Receptor 9)
      0 (Vaccines, Conjugate)
      0 (Vaccines, Subunit)
      25DT549L0G (1018 oligonucleotide)
      9006-59-1 (Ovalbumin)
    • Publication Date:
      Date Created: 20210319 Date Completed: 20211027 Latest Revision: 20211027
    • Publication Date:
      20221213
    • Accession Number:
      PMC7973483
    • Accession Number:
      10.1038/s41598-021-85709-1
    • Accession Number:
      33737644