ArticlePloS one2018
Cell-free synthesis of functional antibody fragments to provide a structural basis for antibody-antigen interaction.
Article in PloS one, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed, 6 citations in OpenAlex.
- Phytochemical Profiling and Computational Screening ofPharmaceuticals (Basel, Switzerland) · 2026Article
- CF2H: a cell-free two-hybrid platform for rapid protein binder screening.Nature communications · 2026Article
- The landscape of antibody production systems: recombinant expression for research, diagnostics and therapy.Frontiers in bioengineering and biotechnology · 2026Review
- Structural insights into endogenous agonist selectivity of aminergic receptors from the octopamine βPNAS nexus · 2025Article
- Analysis of EGFR binding hotspots for design of new EGFR inhibitory biologics.Protein science : a publication of the Protein Society · 2024Review
- Structure-Based Virtual Screening, Molecular Docking, Molecular Dynamics Simulation of EGFR for the Clinical Treatment of Glioblastoma.Applied biochemistry and biotechnology · 2023Article
- Targeting Ras-binding domain of ELMO1 by computational nanobody design.Communications biology · 2023Article
- Potent SARS-CoV-2 neutralizing antibodies with therapeutic effects in two animal models.iScience · 2022Article
- Crystal Structure of an Archaeal Tyrosyl-tRNA Synthetase Bound to Photocaged L-Tyrosine and Its Potential Application to Time-Resolved X-ray Crystallography.International journal of molecular sciences · 2022Article
- Deciphering the language of antibodies using self-supervised learning.Patterns (New York, N.Y.) · 2022Article
- Constructing Cell-Free Expression Systems for Low-Cost Access.ACS synthetic biology · 2022Article
- Article
- Identification of TCR repertoires in functionally competent cytotoxic T cells cross-reactive to SARS-CoV-2.Communications biology · 2021Article
- Crystal structures of HER3 extracellular domain 4 in complex with the designed ankyrin-repeat protein D5.Acta crystallographica. Section F, Structural biology communications · 2021Article
- Current Advancements in Addressing Key Challenges of Therapeutic Antibody Design, Manufacture, and Formulation.Antibodies (Basel, Switzerland) · 2019Review
- Molecular designing, virtual screening and docking study of novel curcumin analogue as mutation (S769L and K846R) selective inhibitor for EGFR.Saudi journal of biological sciences · 2019Article
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Authors and funding
10 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Growing numbers of therapeutic antibodies offer excellent treatment strategies for many diseases. Elucidation of the interaction between a potential therapeutic antibody and its target protein by structural analysis reveals the mechanism of action and offers useful information for developing rational antibody designs for improved affinity. Here, we developed a rapid, high-yield cell-free system using dialysis mode to synthesize antibody fragments for the structural analysis of antibody-antigen complexes. Optimal synthesis conditions of fragments (Fv and Fab) of the anti-EGFR antibody 059-152 were rapidly determined in a day by using a 30-μl-scale unit. The concentration of supplemented disulfide isomerase, DsbC, was critical to obtaining soluble antibody fragments. The optimal conditions were directly applicable to a 9-ml-scale reaction, with linear scalable yields of more than 1 mg/ml. Analyses of purified 059-152-Fv and Fab showed that the cell-free synthesized antibody fragments were disulfide-bridged, with antigen binding activity comparable to that of clinical antibodies. Examination of the crystal structure of cell-free synthesized 059-152-Fv in complex with the extracellular domain of human EGFR revealed that the epitope of 059-152-Fv broadly covers the EGF binding surface on domain III, including residues that formed critical hydrogen bonds with EGF (Asp355EGFR, Gln384EGFR, H409EGFR, and Lys465EGFR), so that the antibody inhibited EGFR activation. We further demonstrated the application of the cell-free system to site-specific integration of non-natural amino acids for antibody engineering, which would expand the availability of therapeutic antibodies based on structural information and rational design. This cell-free system could be an ideal antibody-fragment production platform for functional and structural analysis of potential therapeutic antibodies and for engineered antibody development.
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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.