A5045530041- Innovative Microfluidic and Catalytic Techniques Innovation
- Enzyme Catalysis and Immobilization
- Electrowetting and Microfluidic Technologies
- Chemical Synthesis and Analysis
- Microfluidic and Capillary Electrophoresis Applications
- Biochemical and Structural Characterization
- Analytical Chemistry and Chromatography
- Mass Spectrometry Techniques and Applications
- Scientific Computing and Data Management
- Biosensors and Analytical Detection
- Microbial Natural Products and Biosynthesis
- Microbial Metabolic Engineering and Bioproduction
- Peptidase Inhibition and Analysis
- Microfluidic and Bio-sensing Technologies
- Ubiquitin and proteasome pathways
- Advanced Data Storage Technologies
University of Cambridge
2021-2025
ETH Zurich
2023
Child Trends
2022
Technical University of Munich
2022
Asparaginyl endopeptidases (AEPs) have recently been widely utilized for peptide and protein modification. Labeling is however restricted to termini, severely limiting flexibility scope in creating diverse conjugates as needed therapeutic diagnostic applications. Here, we use genetic code expansion site-specifically modify target proteins with an isopeptide-linked glycylglycine moiety that serves acceptor nucleophile AEP-mediated transpeptidation various probes containing a tripeptidic...
Droplet microfluidics is a valuable method to "beat the odds" in high throughput screening campaigns such as directed evolution, where hits are infrequent and large library sizes required. Absorbance-based sorting expands range of enzyme families that can be subjected droplet by expanding possible assays beyond fluorescence detection. However, absorbance-activated (AADS) currently ∼10-fold slower than typical fluorescence-activated (FADS), meaning that, comparison, larger portion sequence...
Abstract The ability of unevolved amino acid sequences to become biological catalysts was key the emergence life on Earth. However, billions years evolution separate complex modern enzymes from their simpler early ancestors. To probe how can develop new functions, we use ultrahigh-throughput droplet microfluidics screen for phosphoesterase activity amidst a library more than one million based de novo designed 4-helix bundle. Characterization hits revealed that acquisition function involved...
Abstract Engineering enzyme biocatalysts for higher efficiency is key to enabling sustainable, ‘green’ production processes the chemical and pharmaceutical industry. This challenge can be tackled from two angles: by directed evolution, based on labor-intensive experimental testing of variant libraries, or computational methods, where sequence-function data are used predict biocatalyst improvements. Here, we combine both approaches into a two-week workflow, ultra-high throughput screening...
In this report, we detail direct inject mass spectrometry via a Robotically Automated Vacuum Enabled (RAVE) interface that utilizes commercially available capillary electrophoresis hardware to directly samples for (MS) at sampling rate of approximately 12s per sample. This system enables electrospray ionization from standard 48, 96 or 384-well plates with minimal investment in and software, utilizing custom developed web app provides autosampler control analysis raw extracted data the...
Enzyme engineering and discovery are crucial for a sustainable future bioeconomy. Harvesting new biocatalysts from large libraries through directed evolution or functional metagenomics requires accessible, rapid assays. Ultrahigh-throughput screening formats often require optical readouts, leading to the use of model substrates that may misreport target activity necessitate bespoke synthesis. This is particular challenge when glycosyl hydrolases, which leverage molecular recognition beyond...
Droplet microfluidics allows one to address the ever-increasing demand screen large libraries of biological samples. Absorbance spectroscopy complements golden standard fluorescence detection by label free target identification and providing more quantifiable data. However, this is limited speed sensitivity. In paper we increase sorting including acoustofluidics, achieving rates droplets 1 kHz. We improved device design for absorbance using fibre-based interrogation samples with integrated...
Nonribosomal peptides constitute an important class of natural products that display a wide range bioactivities. They are biosynthesized by large assembly lines called nonribosomal peptide synthetases (NRPSs). Engineering NRPS modules represents attractive strategy for generating customized the production variants with improved properties. Here, we explored yeast elongation and termination as high-throughput screening platform assaying adenylation domain activity altering substrate...
Abstract Enzyme engineering and discovery are crucial for a future sustainable bioeconomy, harvesting new biocatalysts from large libraries through directed evolution or functional metagenomics requires accessible, rapid assays. Ultra-high throughput screening can often require an optical readout, leading to the use of model substrates that may not accurately report on activity target reaction bespoke synthesis. In contrast, coupled assays represent modular ‘plug-and-play’ system, where any...
Abstract Droplet microfluidics is a valuable method to ‘beat the odds’ in high throughput screening campaigns such as directed evolution, where hits are infrequent and large library sizes required. Absorbance-based sorting expands landscape of range enzyme families that can be subjected droplet by expanding possible assays beyond fluorescence detection. However, absorbance activated (AADS) currently ∼10-fold slower than typical (FADS), meaning that, comparison, larger portion sequence space...
Abstract Droplet microfluidics allows one to address the ever-increasing demand screen large libraries of biological samples. Absorbance spectroscopy complements golden standard fluorescence detection by label free target identification and providing more quantifiable data. However, this is limited speed sensitivity. In paper we increase sorting including acoustofluidics, achieving rates droplets 1 kHz. We improved devices design for absorbance using fibre-based interrogation samples with...
ABSTRACT The ability of unevolved amino acid sequences to become biological catalysts was key the emergence life on Earth. However, billions years evolution separate complex modern enzymes from their simpler early ancestors. To study how can develop new functions, we screened for enzymatic activity in a collection > 1 million novel based de novo 4-helix bundle library semi-random sequences. mirror evolutionary selection function, using ultrahigh-throughput droplet microfluidics identify...