A5031398219- Monoclonal and Polyclonal Antibodies Research
- Glycosylation and Glycoproteins Research
- Escherichia coli research studies
- Enzyme Structure and Function
- Protein Structure and Dynamics
- Parkinson's Disease Mechanisms and Treatments
- Alzheimer's disease research and treatments
- Neurological disorders and treatments
- Genetic Neurodegenerative Diseases
- Protein purification and stability
- Bacterial Genetics and Biotechnology
- Cellular transport and secretion
- Galectins and Cancer Biology
- Viral Infectious Diseases and Gene Expression in Insects
- Prion Diseases and Protein Misfolding
- Antibiotic Resistance in Bacteria
- Advanced Fluorescence Microscopy Techniques
- RNA and protein synthesis mechanisms
- Biochemical and Structural Characterization
- Proteins in Food Systems
- Synthesis and Properties of Aromatic Compounds
- Enterobacteriaceae and Cronobacter Research
- Nanofabrication and Lithography Techniques
- Silk-based biomaterials and applications
- Brucella: diagnosis, epidemiology, treatment
AstraZeneca (United Kingdom)
2016-2022
University of Cambridge
2009-2021
Integrated Cardio Metabolic Centre
2021
Karolinska Institutet
2021
Vrije Universiteit Brussel
1998-2008
University of Liège
2008
Vlaams Instituut voor Biotechnologie
2002-2006
Institut de génétique et de biologie moléculaire et cellulaire
2005
Inserm
2005
Centre National de la Recherche Scientifique
2005
Clefts on protein surfaces are avoided by antigen-combining sites of conventional antibodies, in contrast to heavy-chain antibodies (HCAbs) camelids that seem be attracted enzymes’ substrate pockets. The explanation for this pronounced preference HCAbs was investigated. Eight single domain antigen-binding fragments (VHH) with nanomolar affinities lysozyme were isolated from three immunized dromedaries. Six eight VHHs compete small inhibitors. This ratio active site binders is also found...
Summary Mannose‐binding type 1 pili are important virulence factors for the establishment of Escherichia coli urinary tract infections (UTIs). These initiated by adhesion uropathogenic E. to uroplakin receptors in uroepithelium via FimH adhesin located at tips pili. Blocking bacterial is able prevent infection. Here, we provide first time binding data molecular events underlying fimbrial adherence, crystallographic analyses receptor domains from a and K‐12 strain, affinity measurements with...
Abstract Naturally spun silks generate fibres with unique properties, including strength, elasticity and biocompatibility. Here we describe a microfluidics-based strategy to spin liquid native silk, obtained directly from the silk gland of Bombyx mori silkworms, into micron-scale capsules controllable geometry variable levels intermolecular β-sheet content in their protein shells. We demonstrate that such micrococoons can store internally otherwise highly unstable for several months without...
Nanobodies are single-domain fragments of camelid antibodies that emerging as versatile tools in biotechnology. We describe here the interactions a specific nanobody, NbSyn87, with monomeric and fibrillar forms α-synuclein (αSyn), 140-residue protein whose aggregation is associated Parkinson's disease. have characterized these using range biophysical techniques, including nuclear magnetic resonance circular dichroism spectroscopy, isothermal titration calorimetry quartz crystal microbalance...
Therapeutics designed to target α-synuclein (α-syn) aggregation may be critical in halting the progression of pathology Parkinson's disease (PD) patients. Nanobodies are single-domain antibody fragments that bind with specificity, but allow readier genetic engineering and delivery. When expressed intracellularly as intrabodies, anti-α-syn nanobodies fused a proteasome-targeting proline, aspartate or glutamate, serine, threonine (PEST) motif can modulate monomeric concentrations proteins....
The aggregation of the amyloid β peptide (Aβ) into fibrils is a defining characteristic Alzheimer's disease. Because complexity this process, effective therapeutic inhibitors will need to target specific microscopic steps that lead production neurotoxic species. We introduce strategy for generating fibril-specific antibodies selectively suppress fibril-dependent secondary nucleation 42-residue form Aβ (Aβ42). step because it has been shown produce majority species during Aβ42. Starting from...
The aggregation of the protein ɑ-synuclein (ɑS) underlies a range increasingly common neurodegenerative disorders including Parkinson's disease. One widely explored therapeutic strategy for these conditions is use antibodies to target aggregated ɑS, although detailed molecular-level mechanism action such species remains elusive. Here, we characterize ɑS in vitro presence two ɑS-specific single-domain (nanobodies), NbSyn2 and NbSyn87, which bind highly accessible C-terminal region ɑS.We show...
Whereas antibodies have demonstrated the ability to mimic various compounds, classic heavy/light-chain may be limited in their applications. First, they tend not bind enzyme active site clefts. Second, size and complexity present problems identifying key elements for binding using these produce clinically valuable compounds. We previously shown how cAb-Lys3, a single variable domain fragment derived from lysozyme-specific camel antibody naturally lacking light chains, overcomes first...
Misfolding, abnormal accumulation, and secretion of α-Synuclein (α-Syn) are closely associated with synucleinopathies, including Parkinson's disease (PD). VH14 is a human single domain intrabody selected against the non-amyloid component (NAC) hydrophobic interaction region α-Syn, which critical for initial aggregation. Using neuronal cell lines, we show that as bifunctional nanobody fused to proteasome targeting signal, VH14PEST can counteract heterologous proteostatic effects mutant α-Syn...
Abstract Amyloid fibrils are a hallmark of range neurodegenerative disorders, including Alzheimer’s and Parkinson’s diseases. A detailed understanding the physico-chemical properties different aggregated forms proteins, their interactions with other compounds diagnostic or therapeutic interest, is crucial for devising effective strategies against such Protein aggregates situated at boundary between soluble insoluble structures, challenging to study because classical biophysical techniques,...
Abstract Local delivery of amyloid beta oligomers from the tip a nanopipette, controlled over cell surface, has been used to deliver physiological picomolar oligomer concentrations primary astrocytes or neurons. Calcium influx was observed when as few 2000 were delivered surface. When dosing stopped intracellular calcium returned basal levels below. prevented by presence in pipette extracellular chaperone clusterin, which is known selectively bind oligomers, and specific nanobody beta. These...
The F17-G adhesin at the tip of flexible F17 fimbriae enterotoxigenic Escherichia coli mediates binding to N-acetyl-beta-D-glucosamine-presenting receptors on microvilli intestinal epithelium ruminants. We report 1.7 A resolution crystal structure lectin domain F17-G, both free and in complex with N-acetylglucosamine. monosaccharide is bound side ellipsoid-shaped protein a conserved site around which all natural variations are clustered. model proposed for interaction between F17-fimbriated...
A single-domain fragment, cAb-HuL22, of a camelid heavy-chain antibody specific for the active site human lysozyme has been generated, and its effects on properties I56T D67H amyloidogenic variants lysozyme, which are associated with form systemic amyloidosis, have investigated by wide range biophysical techniques. Pulse-labeling hydrogen−deuterium exchange experiments monitored mass spectrometry reveal that binding fragment strongly inhibits locally cooperative unfolding restores their...
Affinity maturation of classic antibodies supposedly proceeds through the pre-organization reactive germ line conformational isomer. It is less evident to foresee how this can be accomplished by camelid heavy-chain lacking light chains. Although these are subjected somatic hypermutation, their antigen-binding fragment consists a single domain with restricted flexibility in favor binding energy. An derived from dromedary antibody, cAb-Lys3, accumulated five amino acid substitutions CDR1 and...
We report the effects of interaction two camelid antibody fragments, generally called nanobodies, namely cAb-HuL5 and a stabilized more aggregation-resistant variant cAb-HuL5G obtained by protein engineering, on properties amyloidogenic variants human lysozyme, I56T D67H, whose deposition in vital organs including liver, kidney, spleen is associated with familial non-neuropathic systemic amyloidosis. Both NMR spectroscopy X-ray crystallographic studies reveal that binds to α-domain, one...
A central paradigm in immunology states that successful generation of high affinity antibodies necessitates an immense primary repertoire antigen-combining sites. Much the diversity this is provided by varying one antigen binding loop, created inserting randomly a D (diversity) gene out small pool between V and J genes. It therefore assumed any particular D-encoded region surrounded different regions adopts conformation. We have solved structure two lysozyme-specific variable domains...
Intracellular deposits of α-synuclein in the form Lewy bodies are major hallmarks Parkinson's disease (PD) and a range related neurodegenerative disorders. Post-translational modifications (PTMs) increasingly thought to be modulators its structure, function, degradation toxicity. Among these PTMs, phosphorylation near C-terminus at S129 has emerged as dominant pathogenic modification it is consistently observed occur within brain cerebrospinal fluid (CSF) post-mortem PD patients, level...
Abstract Peptides and proteins have evolved to self‐assemble into supramolecular entities through a set of non‐covalent interactions. Such structures materials provide the functional basis life. Crucially, biomolecular assembly processes can be highly sensitive modulated by environmental conditions, including temperature, light, ionic strength pH, providing inspiration for development new classes responsive based on peptide building blocks. Here, it is shown that stimuli‐responsive...