Multiple Site-Specific Installations ofNε-Monomethyl-L-Lysine into Histone Proteins by Cell-Based and Cell-Free Protein Synthesis

ChemBioChem ◽  
2014 ◽  
Vol 15 (12) ◽  
pp. 1830-1838 ◽  
Author(s):  
Tatsuo Yanagisawa ◽  
Mihoko Takahashi ◽  
Takahito Mukai ◽  
Shin Sato ◽  
Masatoshi Wakamori ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Multiple Site ◽  
Histone Proteins ◽  
Site Specific ◽  
Free Protein ◽  
Cell Free Protein Synthesis

scholarly journals High-yield cell-free protein synthesis for site-specific incorporation of unnatural amino acids at two sites

2012 ◽  
Vol 418 (4) ◽  
pp. 652-656 ◽  
Author(s):  
Kiyoshi Ozawa ◽  
Karin V. Loscha ◽  
Kekini V. Kuppan ◽  
Choy Theng Loh ◽  
Nicholas E. Dixon ◽  
...  
Keyword(s):  
Amino Acids ◽  
Protein Synthesis ◽  
Unnatural Amino Acids ◽  
High Yield ◽  
Site Specific ◽  
Free Protein ◽  
Cell Free Protein Synthesis

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

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Adam M. Weiss ◽  
Jainu Ajit ◽  
Tyler J. Albin ◽  
Neeraj Kapoor ◽  
Shilpa Maroju ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
T Cell ◽  
Antigen Presentation ◽  
Specific Antigen ◽  
T Cell Response ◽  
Site Specific ◽  
Cell Production ◽  
Free Protein ◽  
Cpg Oligodeoxynucleotide ◽  
Cell Free Protein Synthesis

AbstractAntigen-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.

Assessing Site-specific PEGylation of TEM-1 β-lactamase with Cell-free Protein Synthesis and Coarse-grained Simulation

2022 ◽  
Author(s):  
Emily Long Zhao ◽  
Mehran Soltani ◽  
Addison K. Smith ◽  
J. Porter Hunt ◽  
Thomas A. Knotts ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Coarse Grained ◽  
Site Specific ◽  
Free Protein ◽  
Cell Free Protein Synthesis

scholarly journals Site-Specific Cleavage of Ribosomal RNA in Escherichia coli-Based Cell-Free Protein Synthesis Systems

PLoS ONE ◽  
2016 ◽  
Vol 11 (12) ◽  
pp. e0168764 ◽  
Author(s):  
Jurek Failmezger ◽  
Robert Nitschel ◽  
Andrés Sánchez-Kopper ◽  
Michael Kraml ◽  
Martin Siemann-Herzberg
Keyword(s):  
Escherichia Coli ◽  
Protein Synthesis ◽  
Ribosomal Rna ◽  
Site Specific ◽  
Free Protein ◽  
Cell Free Protein Synthesis
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