A cell-free workflow for detecting and characterizing RiPP recognition element-precursor peptide interactions DOI Creative Commons
Derek A. Wong, Zachary M. Shaver, Maria D. Cabezas

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: March 26, 2024

Abstract Post-translational modifications (PTMs) are important for the stability and function of many therapeutic proteins peptides. Current methods studying engineering PTM installing often suffer from low-throughput experimental techniques. Here we describe a generalizable, in vitro workflow coupling cell-free protein synthesis (CFPS) with AlphaLISA rapid expression testing proteins. We apply our to two representative classes peptide therapeutics: ribosomally synthesized post-translationally modified peptides (RiPPs) conjugate vaccines. First, demonstrate how can be used characterize binding activity RiPP recognition elements, an first step biosynthesis, integrated into biodiscovery pipeline computationally predicted products. Then, adapt study engineer oligosaccharyltransferases (OSTs) involved vaccine production, enabling identification mutant OSTs sites within carrier that enable high efficiency production In total, expect will accelerate design-build-test cycles PTMs.

Language: Английский

Glycovaccinology: The design and engineering of carbohydrate-based vaccine components DOI Creative Commons

Sophia W. Hulbert,

Primit Desai,

Michael C. Jewett

et al.

Biotechnology Advances, Journal Year: 2023, Volume and Issue: 68, P. 108234 - 108234

Published: Aug. 7, 2023

Language: Английский

Citations

8
Bacterial glycoengineering: Cell-based and cell-free routes for producing biopharmaceuticals with customized glycosylation DOI

Jaymee A. Palma,

Mehman Bunyatov,

Sophia W. Hulbert

et al.

Current Opinion in Chemical Biology, Journal Year: 2024, Volume and Issue: 81, P. 102500 - 102500

Published: July 10, 2024

Language: Английский

Citations

2
Establishing a Cell-Free Glycoprotein Synthesis System for Enzymatic N-GlcNAcylation DOI
Madison A. DeWinter, Derek A. Wong, Regina Fernández-Morales

et al.

ACS Chemical Biology, Journal Year: 2024, Volume and Issue: 19(7), P. 1570 - 1582

Published: June 27, 2024

-linked glycosylation plays a key role in the efficacy of many therapeutic proteins. One limitation to bacterial glycoengineering human

Language: Английский

Citations

1
Molecular serotyping of diarrheagenic Escherichia coli with a MeltArray assay reveals distinct correlation between serotype and pathotype DOI Creative Commons
Chen Du, Yiqun Liao,

Congcong Ding

et al.

Gut Microbes, Journal Year: 2024, Volume and Issue: 16(1)

Published: Sept. 18, 2024

Diarrheagenic

Language: Английский

Citations

1
A Cell-Free Protein Synthesis Platform to Produce a Clinically Relevant Allergen Panel DOI
Ariel Helms Thames, Clayton H. Rische, Yun Cao

et al.

ACS Synthetic Biology, Journal Year: 2023, Volume and Issue: 12(8), P. 2252 - 2261

Published: Aug. 8, 2023

Allergens are used in the clinical diagnosis (e.g., skin tests) and treatment immunotherapy) of allergic diseases. With growing interest molecular allergy diagnostics precision therapies, new tools needed for producing allergen-based reagents. As a step to address this need, we demonstrate cell-free protein synthesis approach allergen production clinically relevant panel composed common allergens spanning wide range phylogenetic kingdoms. We show that produced with can be recognized by allergen-specific immunoglobulin E (IgE), either monoclonals or patient sera. also expressed activate human cells such as peripheral blood basophils CD34+ progenitor-derived mast an IgE-dependent manner. anticipate platform will enable diagnostic therapeutic technologies, providing useful treatments both allergist patient.

Language: Английский

Citations

2
Advances in Bacterial Oligosaccharyltransferase Structure Elucidation and Potential Application to Glycoconjugate Vaccine Design DOI Creative Commons

Riye Lu,

Pengwei Li,

Li Zhu

et al.

Frontiers in Bioscience-Landmark, Journal Year: 2023, Volume and Issue: 28(11)

Published: Nov. 28, 2023

Glycosylation is one of the most common post-translational modifications proteins across all kingdoms life. Diverse monosaccharides and polysaccharides can be attached to a range amino acid residues generating N-glycosylation, O-glycosylation, C-glycosylation, S-glycosylation, as well P-glycosylation. The functions eukaryotic glycosylation system during protein folding in endoplasmic reticulum (ER) Golgi are well-studied. Increasing evidence recent decade has demonstrated presence oligosaccharyltransferases (OSTs) bacteria archaea. In particular, oligosaccharyltransferase (PglB) Campylobacter jejuni (PglL) enzyme Neisseria meningitidis characterized OSTs that catalyze bacterial N-linked O-linked glycosylation, respectively. Glycoprotein administered glycoconjugate vaccines have been shown effective prophylactic protect against numerous pathogenic bacteria. chemical synthesis glycoproteins complex expensive, which limits its application development vaccines. However, studies biosynthesis realizable by transferring PglB, plasmid encoding substrate protein, or PglL, genes for glycan Escherichia coli. This strategy applied using engineered host E. review summarizes structure mechanism action OSTs, PglB discusses their potential vaccine design.

Language: Английский

Citations

2
A cell-free workflow for detecting and characterizing RiPP recognition element-precursor peptide interactions DOI Creative Commons
Derek A. Wong, Zachary M. Shaver, Maria D. Cabezas

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: March 26, 2024

Abstract Post-translational modifications (PTMs) are important for the stability and function of many therapeutic proteins peptides. Current methods studying engineering PTM installing often suffer from low-throughput experimental techniques. Here we describe a generalizable, in vitro workflow coupling cell-free protein synthesis (CFPS) with AlphaLISA rapid expression testing proteins. We apply our to two representative classes peptide therapeutics: ribosomally synthesized post-translationally modified peptides (RiPPs) conjugate vaccines. First, demonstrate how can be used characterize binding activity RiPP recognition elements, an first step biosynthesis, integrated into biodiscovery pipeline computationally predicted products. Then, adapt study engineer oligosaccharyltransferases (OSTs) involved vaccine production, enabling identification mutant OSTs sites within carrier that enable high efficiency production In total, expect will accelerate design-build-test cycles PTMs.

Language: Английский

Citations

0