ArticlemAbs
Aglycosylated antibodies and antibody fragments produced in a scalable in vitro transcription-translation system.
Article in mAbs. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 64 papers.
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Who cites it
64 citing papers in PubMed.
- Nucleases and Their Inhibitors: Exploring Biological Roles, Industrial Applications, and Challenges in Heterologous Expression.Biotechnology journal · 2026Review
- Phosphoserine as an Alternative Energy Source forACS 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
- BeyondChemical reviews · 2025Review
- Cell-Free Gene Expression: Methods and Applications.Chemical reviews · 2025Review
- Engineering cell-free systems by chemoproteomic-assisted phenotypic screening.RSC chemical biology · 2024Article
- When synthetic biology meets medicine.Life medicine · 2024Review
- At-Home, Cell-Free Synthetic Biology Education Modules for Transcriptional Regulation and Environmental Water Quality Monitoring.ACS synthetic biology · 2023Article
- Enabling technology and core theory of synthetic biology.Science China. Life sciences · 2023Review
- An Integrated In Vivo/In Vitro Protein Production Platform for Site-Specific Antibody Drug Conjugates.Bioengineering (Basel, Switzerland) · 2023Article
- At-home, cell-free synthetic biology education modules for transcriptional regulation and environmental water quality monitoring.bioRxiv : the preprint server for biology · 2023Article
- Development of an E. coli strain for cell-free ADC manufacturing.Biotechnology and bioengineering · 2022Article
- Efficient production of immunologically active Shigella invasion plasmid antigens IpaB and IpaH using a cell-free expression system.Applied microbiology and biotechnology · 2022Article
- The Impact of Glycoengineering on the Endoplasmic Reticulum Quality Control System in Yeasts.Frontiers in molecular biosciences · 2022Review
- Simple Extract Preparation Methods for E. coli-Based Cell-Free Expression.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Optimization of E. Coli Tip-Sonication for High-Yield Cell-Free Extract using Finite Element Modeling.AIChE journal. American Institute of Chemical Engineers · 2021Article
- A lysate proteome engineering strategy for enhancing cell-free metabolite production.Metabolic engineering communications · 2021Article
- Optimising protein synthesis in cell-free systems, a review.Engineering biology · 2021Article
- Biofoundry-assisted expression and characterization of plant proteins.Synthetic biology (Oxford, England) · 2021Article
4 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
13 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We describe protein synthesis, folding and assembly of antibody fragments and full-length aglycosylated antibodies using an Escherichia coli-based open cell-free synthesis (OCFS) system. We use DNA template design and high throughput screening at microliter scale to rapidly optimize production of single-chain Fv (scFv) and Fab antibody fragments that bind to human IL-23 and IL-13α1R, respectively. In addition we demonstrate production of aglycosylated immunoglobulin G (IgG 1) trastuzumab. These antibodies are produced rapidly over several hours in batch mode in standard bioreactors with linear scalable yields of hundreds of milligrams/L over a 1 million-fold change in scales up to pilot scale production. We demonstrate protein expression optimization of translation initiation region (TIR) libraries from gene synthesized linear DNA templates, optimization of the temporal assembly of a Fab from independent heavy chain and light chain plasmids and optimized expression of fully assembled trastuzumab that is equivalent to mammalian expressed material in biophysical and affinity based assays. These results illustrate how the open nature of the cell-free system can be used as a seamless antibody engineering platform from discovery to preclinical development of aglycosylated monoclonal antibodies and antibody fragments as potential therapeutics.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.