ArticleMicrobial cell factories2016
A nanobody:GFP bacterial platform that enables functional enzyme display and easy quantification of display capacity.
Article in Microbial cell factories, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed.
- Engineered bacteria as an orally administered anti-viral treatment and immunization system.Gut microbes · 2025Article
- Resurfacing promotes antibacterial activity of a lipid A-binding nanobody.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Bacterial Systems as a Precision Delivery Platform of Therapeutic Peptides for Cancer Therapy.Polymer science & technology (Washington, D.C.) · 2025Review
- Genetically surface-modifiedBiotechnology reports (Amsterdam, Netherlands) · 2024Article
- A New Bacterial Chassis for Enhanced Surface Display of Recombinant Proteins.Cellular and molecular bioengineering · 2024Article
- Quantitative measurement of cell-surface displayed proteins based on split-GFP assembly.Microbial cell factories · 2024Article
- Research Progresses and Applications of Fluorescent Protein Antibodies: A Review Focusing on Nanobodies.International journal of molecular sciences · 2023Review
- Direct selection of functional fluorescent-protein antibody fusions by yeast display.PloS one · 2023Article
- Nanoscale Features of Tunable Bacterial Outer Membrane Models Revealed by Correlative Microscopy.Langmuir : the ACS journal of surfaces and colloids · 2022Article
- In vitro inhibition of cancer angiogenesis and migration by a nanobody that targets the orphan receptor Tie1.Cellular and molecular life sciences : CMLS · 2022Article
- Surface display as a functional screening platform for detecting enzymes active on PET.Microbial cell factories · 2021Article
- Exploring cellular biochemistry with nanobodies.The Journal of biological chemistry · 2020Review
- Bacterial Cell Display as a Robust and Versatile Platform for Engineering Low-Affinity Ligands and Enzymes.Chembiochem : a European journal of chemical biology · 2020Article
- Nanobody-Based Probes for Subcellular Protein Identification and Visualization.Frontiers in cellular neuroscience · 2020Review
- An improved yeast surface display platform for the screening of nanobody immune libraries.Scientific reports · 2019Article
Corrections and comments
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Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundBacterial surface display is an attractive technique for the production of cell-anchored, functional proteins and engineering of whole-cell catalysts. Although various outer membrane proteins have been used for surface display, an easy and versatile high-throughput-compatible assay for evaluating and developing surface display systems is missing.
resultsUsing a single domain antibody (also called nanobody) with high affinity for green fluorescent protein (GFP), we constructed a system that allows for fast, fluorescence-based detection of displayed proteins. The outer membrane hybrid protein LppOmpA and the autotransporter C-IgAP exposed the nanobody on the surface of Escherichia coli with very different efficiency. Both anchors were capable of functionally displaying the enzyme Chitinase A as a fusion with the nanobody, and this considerably increased expression levels compared to displaying the nanobody alone. We used flow cytometry to analyse display capability on single-cell versus population level and found that the signal peptide of the anchor has great effect on display efficiency.
conclusionsWe have developed an inexpensive and easy read-out assay for surface display using nanobody:GFP interactions. The assay is compatible with the most common fluorescence detection methods, including multi-well plate whole-cell fluorescence detection, SDS-PAGE in-gel fluorescence, microscopy and flow cytometry. We anticipate that the platform will facilitate future in-depth studies on the mechanism of protein transport to the surface of living cells, as well as the optimisation of applications in industrial biotech.
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