Scalable Cell‐Free Production of Active T7 RNA Polymerase DOI

R. Rezvani,

Rochelle Aw, J. Chan

et al.

Biotechnology and Bioengineering, Journal Year: 2025, Volume and Issue: unknown

Published: April 29, 2025

ABSTRACT The SARS‐CoV‐2 pandemic highlighted the urgent need for biomanufacturing paradigms that are robust and fast. Here, we demonstrate rapid process development scalable cell‐free production of T7 RNA polymerase, a critical component in mRNA vaccine synthesis. We carry out 1‐L gene expression (CFE) reaction achieves over 90% purity, low endotoxin levels, enhanced activity relative to commercial polymerase. To achieve this demonstration, implement rolling circle amplification circumvent difficulties DNA template generation, tune conditions, such as temperature, additives, purification tags, agitation, boost yields. similar quality titer polymerase more than four orders magnitude volume. This proof principle positions CFE viable solution decentralized biotherapeutic manufacturing, enhancing preparedness future public health crises or emergent threats.

Language: Английский

Cell-free protein synthesis platforms for accelerating drug discovery DOI Open Access
Filippo Caschera

Biotechnology Notes, Journal Year: 2025, Volume and Issue: 6, P. 126 - 132

Published: Jan. 1, 2025

Language: Английский

Citations

0
Stability insights into synthetic biology enabled biosensors: A case study with SARS-CoV2 toehold-based colorimetric sensor DOI Creative Commons

Swetha Mariam Stanley,

Harvinder Kour Khera

Sensing and Bio-Sensing Research, Journal Year: 2025, Volume and Issue: unknown, P. 100775 - 100775

Published: March 1, 2025

Language: Английский

Citations

0
Cell-Free Expression of Soluble Leafhopper Proteins from Brochosomes DOI

Caleb G. Lay,

Gabriel Burks, Zheng Li

et al.

ACS Synthetic Biology, Journal Year: 2025, Volume and Issue: unknown

Published: March 7, 2025

Brochosomes are proteinaceous nanostructures produced by leafhopper insects with superhydrophobic and antireflective properties. Unfortunately, the production study of brochosome-based materials has been limited poor understanding their major constituent subunit proteins, known as brochosomins, well sensitivity to redox conditions due essential disulfide bonds. Here, we used cell-free gene expression (CFE) achieve recombinant analysis brochosomin proteins. Through optimization environment, reaction temperature, bond isomerase concentration, achieved soluble yields up 341 ± 30 μg/mL. Analysis using dynamic light scattering transmission electron microscopy revealed distinct aggregation patterns among mixtures different expressed brochosomins. We anticipate that CFE methods developed here will accelerate ability change geometries properties natural modified brochosomes, facilitate structural other poorly understood protein complexes.

Language: Английский

Citations

0
Exozymes for Biomanufacturing: Toward Clarity and Precision in the Cell-Free Space DOI
Adrian Brückner, Bastian Vögeli, John M. Billingsley

et al.

GEN Biotechnology, Journal Year: 2025, Volume and Issue: unknown

Published: April 14, 2025

Language: Английский

Citations

0
Scalable Cell‐Free Production of Active T7 RNA Polymerase DOI

R. Rezvani,

Rochelle Aw, J. Chan

et al.

Biotechnology and Bioengineering, Journal Year: 2025, Volume and Issue: unknown

Published: April 29, 2025

ABSTRACT The SARS‐CoV‐2 pandemic highlighted the urgent need for biomanufacturing paradigms that are robust and fast. Here, we demonstrate rapid process development scalable cell‐free production of T7 RNA polymerase, a critical component in mRNA vaccine synthesis. We carry out 1‐L gene expression (CFE) reaction achieves over 90% purity, low endotoxin levels, enhanced activity relative to commercial polymerase. To achieve this demonstration, implement rolling circle amplification circumvent difficulties DNA template generation, tune conditions, such as temperature, additives, purification tags, agitation, boost yields. similar quality titer polymerase more than four orders magnitude volume. This proof principle positions CFE viable solution decentralized biotherapeutic manufacturing, enhancing preparedness future public health crises or emergent threats.

Language: Английский

Citations

0