ArticleScientific reports2019
Constructive approach for synthesis of a functional IgG using a reconstituted cell-free protein synthesis system.
Article in Scientific reports, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed.
- Impact of Reducing Agents on Protein Synthesis in a Reconstituted Cell-Free Protein Synthesis System.ACS synthetic biology · 2026Article
- Design-driven optimization of low-cost reagent formulations for reproducible and high-yielding cell-free gene expression.Nature communications · 2026Article
- The landscape of antibody production systems: recombinant expression for research, diagnostics and therapy.Frontiers in bioengineering and biotechnology · 2026Review
- Parameterization of cell-free systems with time-series data using KETCHUP.PLoS computational biology · 2025Article
- Automated and Programmable Cell-Free Systems for Scalable Synthetic Biology with a Focus on Biofoundry Integration.Journal of microbiology and biotechnology · 2025Review
- Developing anACS synthetic biology · 2025Article
- FASTIA: A rapid and accessible platform for protein variant interaction analysis demonstrated with a single-domain antibody.Protein science : a publication of the Protein Society · 2025Article
- Cell-Free Gene Expression: Methods and Applications.Chemical reviews · 2025Review
- Review
- N-Terminal Amino Acid Affects the Translation Efficiency at Lower Temperatures in a Reconstituted Protein Synthesis System.International journal of molecular sciences · 2024Article
- Regulated N-Terminal Modification of Proteins Synthesized Using a Reconstituted Cell-Free Protein Synthesis System.ACS synthetic biology · 2023Article
- A rapid cell-free expression and screening platform for antibody discovery.Nature communications · 2023Article
- Synthesis of an Anti-CD7 Recombinant Immunotoxin Based on PE24 in CHO andInternational journal of molecular sciences · 2022Article
- Cell-Free Protein Synthesis for the Screening of Novel Azoreductases and Their Preferred Electron Donor.Chembiochem : a European journal of chemical biology · 2022Article
- A Spider Toxin Exemplifies the Promises and Pitfalls of Cell-Free Protein Production for Venom Biodiscovery.Toxins · 2021Article
- A Relationship between NTP and Cell Extract Concentration for Cell-Free Protein Expression.Life (Basel, Switzerland) · 2021Article
- Review
- Tuning the Cell-Free Protein Synthesis System for Biomanufacturing of Monomeric Human Filaggrin.Frontiers in bioengineering and biotechnology · 2020Article
- Methods to reduce variability inSynthetic and systems biotechnology · 2019Article
- Review
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Authors and funding
3 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
IgG is an indispensable biological experimental tool as well as a widely-used therapeutic protein. However, cell culture-based expression of monoclonal IgG is costly and time-consuming, making this process difficult to use for high-throughput screening in early-stage evaluation of biologics. With the goal of establishing a fast, simple, and robust high-throughput expression system for IgG, we implemented the synthesis of functional aglycosylated IgG by constructive approach based on a reconstituted prokaryotic cell-free protein synthesis system (PURE system). Optimization of the PURE system revealed that the following factors and reaction conditions were needed for IgG synthesis: (1) inclusion of the disulfide bond isomerase DsbC, (2) adjustment of the GSH/GSSG ratio, (3) inclusion of the molecular chaperone DnaK and its cofactors, and (4) use of an extended incubation time. Synthesis temperature and template DNA ratio (light chain-/heavy chain-encoding) also had been optimized for each IgG. Under optimal conditions, peak production of the anti-HER2 antibody trastuzumab reached 124 µg/mL. Furthermore, the active forms of other IgGs, including IgG1, IgG2, and IgG4 subclasses, also were synthesized. These results provide basic information for the development of novel high-throughput expression and functional screening systems for IgG, as well as useful information for understanding the IgG synthesis process.
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