A5027661645- Tuberculosis Research and Epidemiology
- Mycobacterium research and diagnosis
- Monoclonal and Polyclonal Antibodies Research
- Biochemical and Molecular Research
- Cancer therapeutics and mechanisms
- Pharmacogenetics and Drug Metabolism
- Computational Drug Discovery Methods
- Chemical Reactions and Isotopes
- CAR-T cell therapy research
- RNA and protein synthesis mechanisms
- Bacteriophages and microbial interactions
- Protein purification and stability
- Analytical Chemistry and Chromatography
- vaccines and immunoinformatics approaches
- Protein Structure and Dynamics
- SARS-CoV-2 and COVID-19 Research
- Lymphoma Diagnosis and Treatment
- Crystallization and Solubility Studies
- Enzyme Structure and Function
- Nanofabrication and Lithography Techniques
- Viral Infectious Diseases and Gene Expression in Insects
- 14-3-3 protein interactions
- Bacterial Genetics and Biotechnology
- CRISPR and Genetic Engineering
- Advanced Biosensing Techniques and Applications
University of Cambridge
2009-2017
Max Planck Institute of Biophysics
2006-2012
Max Planck Society
2012
Abstract Peripheral T cell lymphomas are typically aggressive with a poor prognosis. Unlike other hematologic malignancies, the lack of target antigens to discriminate healthy from malignant cells limits efficacy immunotherapeutic approaches. The receptor expresses one two highly homologous chains [T β-chain constant (TRBC) domains 1 and 2] in mutually exclusive manner, making it promising target. Here we demonstrate specificity redirection by rational design using structure-guided...
The early phase of protein drug development has traditionally focused on target binding properties leading to a desired mode therapeutic action. As more therapeutics pass through the pipeline; however, it is clear that non-optimal biophysical can emerge, particularly as proteins are formulated at high concentrations, causing aggregation or polyreactivity. Such late-stage "developability" problems lead delay failure in traversing process. Aggregation propensity also correlated with increased...
Pieces of the puzzle: The first fragment-based approach was used to target cytochrome P450 enzymes (CYPs) for drug development (see scheme). experiments provide new insights into binding site essential Mycobacterium tuberculosis CYP121 enzyme, and resulted in a promising novel lead compound based on fragment merging. Detailed facts importance specialist readers are published as "Supporting Information". Such documents peer-reviewed, but not copy-edited or typeset. They made available...
The essential enzyme CYP121 is a target for drug development against antibiotic resistant strains of Mycobacterium tuberculosis. A triazol-1-yl phenol fragment 1 was identified to bind using cascade biophysical assays. Synthetic merging and optimization produced 100-fold improvement in binding affinity, yielding lead compound 2 (KD = 15 μM). Deconstruction into its component retrofragments allowed the group efficiency structural motifs be assessed, identification more LE scaffolds...
The construction of large libraries in mammalian cells allows the direct screening millions molecular variants for binding properties a cell type relevant or production. We have created up to 10 million clones displaying repertoire IgG-formatted antibodies on surface. TALE nucleases CRISPR/Cas9 were used integration antibody genes into single genomic locus, thereby rapidly achieving stable expression and transcriptional normalization. utility system is illustrated by affinity maturation...
With the ever-increasing instances of resistance to frontline TB drugs there is need develop novel strategies fight worldwide epidemic.
Membrane proteins comprise up to one-third of prokaryotic and eukaryotic genomes, but only a very small number membrane protein structures are known. challenging targets for structural biology, primarily due the difficulty in producing purifying milligram quantities these proteins. We evaluating different methods produce purify large numbers subsequent functional analysis. Here, we present comparative expression data 37 target proteins, all them secondary transporters, from mesophilic...
Passive immunization using monoclonal antibodies will play a vital role in the fight against COVID-19. The recent emergence of viral variants with reduced sensitivity to some current and vaccines highlights importance broad cross-reactivity. This study describes deep-mining antibody repertoires hospitalized COVID-19 patients phage display technology B cell receptor (BCR) repertoire sequencing isolate neutralizing gain insights into early response. comprehensive discovery approach has yielded...
A structure-guided fragment-based approach was used to target the lipophilic allosteric binding site of Mycobacterium tuberculosis EthR. This elongated channel has many hydrophobic residues lining site, with few opportunities for hydrogen bonding. We demonstrate that a involving inclusion flexible fragments in library leads an efficient exploration chemical space, fragment can lead extension cavity, and are able identify hydrogen-bonding this environment not exploited Nature. In present...
Freedom to merge: A combination of crystal structure examination and in silico predictions made it possible overcome the conformational limitations fragment merging escape internal strain a series weakly binding merged fragments that target M. tuberculosis CYP121. The insights attained provide new perspective guide for prioritizing synthetic efforts toward future ongoing fragment-based ligand discovery campaigns.
Small-molecule inhibitors of the mycobacterial transcriptional repressor EthR have previously been shown to act as boosters second-line antituberculosis drug ethionamide. Fragment-based discovery approaches used in past make highly potent with ethionamide boosting activity both vitro and ex vivo. Herein, we report development fragment-sized ligands nanomolar minimum effective concentration values for Mycobacterium tuberculosis whole-cell assays.
Thymidylate synthase X (ThyX) represents an attractive target for tuberculosis drug discovery. Herein, we selected 16 compounds through a virtual screening approach. We solved the first X-ray crystal structure of Thermatoga maritima (Tm) ThyX in complex with nonsubstrate analog inhibitor. Given active site similarities between Mycobacterium (Mtb-ThyX) and Tm-ThyX, our paves way structure-based design novel antimycobacterial compounds. The 1H-imidazo[4,5-d]pyridazine was identified as...
A new strategy is proposed for batch crystallization of proteins in solution-growth or gel-growth by using the method inside capillary tubes applying magnetic fields. Four with differing proportions α-helices and β-sheets crystallized five different crystallographic space groups are studied, allowing an analysis anisotropy diamagnetic susceptibility peptide bond as well polarity presence a strong field 11.75 T. The crystal quality shown to be improved orient protein molecules, (high...
The use of an organic solvent-based gel prepared from polyethylene oxide and a polyvinyl alcohol hydrogel for protein crystallization was investigated. preparation, properties application the gels are described, advantages limitations approach discussed. compared with agar, which is popular aqueous used crystallization. growth behaviour diffraction quality crystals in these media were evaluated two model soluble proteins, thaumatin lysozyme, bacterial membrane TolC AcrB.
Abstract Developing therapeutic antibodies is a challenging endeavour, often requiring large-scale screening to produce initial binders, that still require optimisation for developability. We present computational pipeline the discovery and design of antibody candidates, which incorporates physics- AI-based methods generation, assessment, validation developable candidate against diverse epitopes, via efficient few-shot experimental screens. demonstrate these orthogonal can lead promising...
Abstract The members of the cation diffusion facilitator (CDF) family transport heavy metal ions and play an important function in zinc ion homeostasis cell. A recent structure Escherichia coli CDF transporter protein YiiP has revealed its dimeric nature autoregulatory mechanism. Here, we report cloning heterologous production four different transporters, two each from pathogenic mesophilic bacterium Salmonella typhimurium hyperthermophilic Aquifex aeolicus , E. host cells. STM0758 S. was...
Teile des Puzzles: Ein fragmentbasierter Ansatz wurde erstmals zur Untersuchung von Cytochrom-P450-Enzymen (CYPs) in der Wirkstoffentwicklung eingesetzt (siehe Schema). Die Experimente bieten neue Einblicke die Bindungsstellen CYP121-Enzyms aus Mycobacterium tuberculosis und führten zu einer vielversprechenden neuen Leitverbindung. Detailed facts of importance to specialist readers are published as ”Supporting Information”. Such documents peer-reviewed, but not copy-edited or typeset. They...
Abstract Peripheral T cell lymphomas are typically aggressive with a poor prognosis. Unlike other hematologic malignancies, the lack of target antigens to discriminate healthy from malignant cells has limited efficacy immunotherapeutic approaches. The receptor expresses one two highly homologous chains [T β-chain constant (TRBC) domains 1 and 2] in mutually exclusive manner, making it promising target. We previously described an antibody unique TRBC1 specificity (Jovi-1). Here we demonstrate...
Abstract Passive immunisation using monoclonal antibodies will play a vital role in the fight against COVID-19. Until now, majority of anti-SARS-CoV-2 antibody discovery efforts have relied on screening B cells patients convalescent phase. Here, we describe deep-mining repertoires hospitalised COVID-19 combination phage display technology and cell receptor (BCR) repertoire sequencing to isolate neutralising gain insights into early response. This comprehensive approach has yielded potent...