F. Hollfelder

Thermostable in vitro transcription-translation for enzyme screening in microdroplets

This paper develops a thermostable in vitro transcription-translation system using Thermus thermophilus extracts for enzyme screening in microdroplets.

Fluorogenic, sub-single-turnover monitoring of enzymatic reactions involving NAD(P)H provides a generalised platform for directed ultrahigh throughput evolution of biocatalysts in microdroplets

The paper develops a fluorogenic assay for NAD(P)H detection to enable high-throughput screening of biocatalysts in microdroplets, demonstrating improved sensitivity and applicability for glycosyl hydrolases and IREDs.

Ultrahigh throughput evolution of tryptophan synthase in droplets via an aptamer-biosensor

This paper develops a DNA aptamer biosensor to enable high-throughput screening of tryptophan synthase variants in droplets, identifying 5-fold improved enzymes.

Rapid quantitative assessment of small molecule leakage from microdroplets by flow cytometry and improvement of fluorophore retention in biochemical assays

The paper develops a flow cytometry method to quantify small molecule leakage from microdroplets and improves fluorophore retention in biochemical assays.

Versatile Product Detection via Coupled Assays for Ultra-high-throughput Screening of Carbohydrate-Active-Enzymes in Microfluidic Droplets

This paper develops microfluidic droplet assays to enable ultra-high-throughput screening of carbohydrate-active enzymes by detecting unlabelled carbohydrate products through optical signals.

The biosynthesis, degradation, and function of cell wall β-xylosylated xyloglucan mirrors that of arabinoxyloglucan

The paper identifies enzymes involved in the biosynthesis and degradation of β-xylosylated xyloglucan and shows their potential role in gut bacterial nutrition.

Selection of a Promiscuous Minimalist cAMP Phosphodiesterase from a Library of De Novo Designed Proteins

The paper screens a library of de novo designed proteins to identify a promiscuous cAMP phosphodiesterase, finding that truncation and structural dynamics enable catalytic activity.

Microfluidics-enabled fluorescence-activated cell sorting of single pathogen-specific antibody secreting cells for the rapid discovery of monoclonal antibodies

This paper presents a microfluidics-based method to rapidly discover monoclonal antibodies from antigen-specific antibody secreting cells, achieving high-affinity and neutralizing antibodies against SARS-CoV-2.

Ultrahigh-throughput Absorbance Activated Droplet Sorting (UHT-AADS) for enzyme screening at kilohertz frequencies

This paper introduces UHT-AADS, a method that improves absorbance-based droplet sorting speed to match fluorescence sorting, enhancing enzyme screening throughput.

Sorting of droplets at kHz rates using absorbance activated acoustic sorting

This paper introduces an acoustic activated absorbance sorter that increases sorting speed to 1 kHz and improves signal quality by reducing scattering and refraction artefacts.

High throughput steady-state enzyme kinetics measured in a parallel droplet generation and absorbance detection platform

This paper introduces a high-throughput droplet-based system for measuring enzyme kinetics with improved throughput and data quality compared to existing methods.

Functional metagenomic screening in microfluidic droplets identifies a β-glucuronidase in an unprecedented sequence neighbourhood

This study identifies a novel β-glucuronidase from a metagenomic library using microfluidic droplet screening, revealing a new GH3 family member with unique functional and structural properties.

Ultrahigh-throughput directed evolution of a metal-free α/β-hydrolase with a Cys-His-Asp triad into an efficient phosphotriesterase

This paper evolves a metal-free α/β-hydrolase with a Cys-His-Asp triad into an efficient phosphotriesterase using ultrahigh-throughput screening, achieving catalytic efficiencies comparable to metal-dependent enzymes.

Improved RAD51 binders through motif shuffling based on the modularity of BRC repeats

The paper creates and tests chimeric BRC repeats to improve RAD51 binding, finding that shuffled modules can enhance binding affinity and function in human cells.

In vitro Evolution of Antibody Affinity via Insertional Mutagenesis Scanning of an Entire Antibody Variable Region

This paper demonstrates that insertional mutagenesis can enhance antibody affinity, achieving a 256-fold improvement in an anti-IL-13 antibody through InDel mutagenesis combined with point mutations.

Microfluidic platform for live cell imaging of 3D cultures with clone retrieval

The paper presents a microfluidic platform for live cell imaging of 3D cultures with the ability to retrieve individual cells for functional assays.

Access to unexplored regions of sequence space in directed enzyme evolution via insertion/deletion mutagenesis

This study introduces TRIAD, a transposon mutagenesis method to generate InDels for directed enzyme evolution, showing distinct functional outcomes compared to point mutagenesis.

High-Throughput, Lysis-free Screening for sulfatase Activity Using Escherichia coli Autodisplay in Microdroplets

This paper develops a high-throughput, lysis-free screening method using E. coli autodisplay and microdroplet sorting to identify sulfatase variants with improved activity.

Transition State Interactions in a Promiscuous Enzyme: Sulfate and Phosphate Monoester Hydrolysis by Pseudomonas aeruginosa Arylsulfatase

This study investigates the mechanism of sulfate and phosphate monoester hydrolysis by Pseudomonas aeruginosa arylsulfatase using LFERs and KIEs, revealing key roles for active site residues in leaving group stabilization and mechanism shifts.