A5062741806- Click Chemistry and Applications
- Monoclonal and Polyclonal Antibodies Research
- Chemical Synthesis and Analysis
- Biochemical and Structural Characterization
- CRISPR and Genetic Engineering
- Peptidase Inhibition and Analysis
- Enzyme Catalysis and Immobilization
- Biotin and Related Studies
- Advanced biosensing and bioanalysis techniques
- Enzyme Production and Characterization
- Venomous Animal Envenomation and Studies
- RNA and protein synthesis mechanisms
- Viral Infectious Diseases and Gene Expression in Insects
- Molecular Junctions and Nanostructures
- Transgenic Plants and Applications
- Advanced Biosensing Techniques and Applications
- Rabies epidemiology and control
- Glycosylation and Glycoproteins Research
- Polymer Surface Interaction Studies
- Virus-based gene therapy research
- RNA regulation and disease
- Microfluidic and Capillary Electrophoresis Applications
- Protein purification and stability
- Innovative Microfluidic and Catalytic Techniques Innovation
- Cancer Research and Treatments
Novo Nordisk Foundation
2020-2025
Technical University of Denmark
2020-2025
Danmarks Nationalbank
2021
Foundation Center
2021
University of Copenhagen
2014-2020
Radboud University Nijmegen
2008-2014
Radboud Institute for Molecular Life Sciences
2008
Radboud University Medical Center
2008
A strained aza-dibenzocyclooctyne was prepared via a high-yielding synthetic route. Copper-free, strain-promoted click reaction with azides showed excellent kinetics, and functionalised aza-cyclooctyne applied in fast efficient PEGylation of enzymes.
In living cells enzymes catalyze a wide variety of metabolic processes, which involve multiple reaction steps. Efficient transfer the intermediates from one catalytic site to other is achieved by formation macromolecular enzyme complexes. This phenomenon, called channeling, has inspired researchers bring biocatalysts together in an artificial way as well. review describes various vitro strategies have been exploited so far. A distinction made based on degree control over assembly process....
Abstract Host cell proteins (HCPs) are process-related impurities generated during biotherapeutic protein production. HCPs can be problematic if they pose a significant metabolic demand, degrade product quality, or contaminate the final product. Here, we present an effort to create “clean” Chinese hamster ovary (CHO) by disrupting multiple genes eliminate HCPs. Using model of CHO secretion, predict that elimination unnecessary could have non-negligible impact on We analyze HCP content...
Abstract Snakebite envenoming continues to claim many lives across the globe, necessitating development of improved therapies. To this end, broadly-neutralizing human monoclonal antibodies may possess advantages over current plasma-derived antivenoms by offering superior safety and high neutralization capacity. Here, we report establishment a pipeline based on phage display technology for discovery optimization affinity antibodies. This approach yielded recombinant antibody with capacities...
Abstract Oligoclonal mixtures of broadly-neutralizing antibodies can neutralize complex compositions similar and dissimilar antigens, making them versatile tools for the treatment e.g., infectious diseases animal envenomations. However, these biotherapeutics are complicated to develop due their nature. In this work, we describe application various strategies discovery cross-neutralizing nanobodies against key toxins in coral snake venoms using phage display technology. We prepare two...
Abstract Improved therapies are needed against snakebite envenoming, which kills and permanently disables thousands of people each year. Recently developed neutralizing monoclonal antibodies several snake toxins have shown promise in preclinical rodent models. Here, we use phage display technology to discover a human antibody show that this causes antibody-dependent enhancement toxicity (ADET) myotoxin II from the venomous pit viper, Bothrops asper , mouse model envenoming mimics snakebite....
Fluorobenzene probes for protein profiling through selective cysteine labeling have been developed by rational reactivity tuning. Tuning was achieved selecting an electron-withdrawing para substituent in combination with variation of the number fluorine substituents. Optimized chemoselectively arylated residues proteins under aqueous conditions. Probes linked to azide, biotin, or a fluorophore were applicable eGFP and albumin. Selective inhibition proteases also demonstrated probes....
<ns4:p>The properties of biosensors, biomedical implants, and other materials based on immobilized proteins greatly depend the method employed to couple protein molecules their solid support. Covalent, site-specific immobilization strategies are robust can provide level control that is desired in this kind application. Recent advances include use enzymes, such as sortase A, a manner microbeads, glass, hydrogels. Also, self-labeling tags SNAP-tag be employed. Last but not least, chemical...
Abstract A block copolymer was designed to functionalise the surface of polystyrene‐based polymersomes via coaggregation. An α , ω ‐diacetylene‐functionalised poly(ethylene glycol) (PEG) coupled an azide‐terminated polystyrene a Cu(I)‐catalysed cycloaddition produce PS‐ b ‐PEG polymer with acetylene at its hydrophilic extremity. Incorporation this ‘anchor’ compound in bilayer polymersome places bio‐orthogonal group aggregate. Its accessibility demonstrated using azido‐functionalised Candida...
In order to modify proteins in a controlled way, new functionalities need be introduced defined manner. One way accomplish this is by the incorporation of non-natural amino acid which side chain can selectively reacted other molecules. We have investigated whether relatively simple method residue-specific replacement methionine azidohomoalanine used achieve monofunctionalization model enzyme Candida antarctica lipase B. A protein variant was engineered with one additional residue. Due high...
Conversion of amines into azides using imidazole-1-sulfonyl azide as a diazotransfer reagent has proven to be straightforward way introduce targetable handles proteins. We explored whether less toxic and milder conditions than described before could applied. It was shown that the aqueous proceeds without adding Cu(II) not only at pH 11 but also 8.5. The selective towards single amine.
Methods for site-selective chemistry on proteins are in high demand the synthesis of chemically modified biopharmaceuticals, as well applications chemical biology, biosensors and more. Inadvertent N-terminal gluconoylation has been reported during expression with an His tag. Here we report development this side-reaction into a general method highly selective acylation to introduce functional groups. We identify optimized sequence, GHHHn- reaction gluconolactone 4-methoxyphenyl esters...
A human epidermal growth factor receptor 2 (HER2)-specific nanobody called 2Rs15d, containing a His3LysHis6 segment at the C-terminus, was recombinantly produced. Subsequent site-selective acylation on C-terminally activated lysine residue allowed installation of cytotoxin monomethyl auristatin E-functionalized cathepsin B-sensitive payload to provide highly homogenous nanobody–drug conjugate (NBC), which demonstrated high potency and selectivity for HER2-positive breast cancer models.
An effective vaccine against hepatitis C virus (HCV) must elicit the production of broadly neutralizing antibodies (bnAbs) reproducibly E1E2 glycoprotein complex. Little is known about how glycan content affects this process. Ideally, glycans would maximize epitope exposure without compromising antigen stability or exposing new epitopes. However, typical recombinant vaccines contain considerable heterogeneity in content, which can affect antibody response and neutralization potency. Here we...
A three-enzyme cascade reaction was successfully realized in a continuous flow microreactor. The first enzyme (Candida antarctica lipase B, also known as Pseudozyma B) and the third (horseradish peroxidase) of process were immobilized mild non-contact manner via ssDNA-ssDNA interaction discrete zones on capillary wall, whereas second (glucose oxidase) kept mobile phase. unique combined feature patterning, possibility loading stripping, modularity fused silica microchannel is demonstrated. By...
A series of stimulus-responsive elastin-like polypeptide-poly(ethylene glycol) (ELP-PEG) block copolymers was synthesized. The polymeric building blocks were conjugated via the efficient and specific strain-promoted alkyne-azide cycloaddition (SPAAC). For this purpose, ELP PEG functionalized with azide cyclooctyne moieties, respectively. Azides introduced by applying a recently developed pH-controlled diazotransfer reaction on primary amines present in (N-terminus lysine side chains). By...
Elastin-like polypeptides (ELPs) functionalized with azide or alkyne groups were produced biosynthetically and coupled via the Cu-catalyzed azide–alkyne cycloaddition to a variety of (bio)molecules.
Inspired by the multienzyme complexes occurring in nature, enzymes have been brought together vitro as well. We report a co-localization strategy milder than nonspecific cross-linking, and free of any scaffold affinity tags. Using non-natural amino acid incorporation, two heterobifunctional linkers, strain-promoted azide-alkyne cycloaddition conjugation reaction, three metabolic are linked controlled manner. Conjugate formation was demonstrated size-exclusion chromatography gel...
Glycosylated biopharmaceuticals are important in the global pharmaceutical market. Despite importance of their glycan structures, our limited knowledge glycosylation machinery still hinders controllability this critical quality attribute. To facilitate discovery glycosyltransferase specificity and predict glycoengineering efforts, here we extend approach to model N-linked protein as a Markov process. Our leverages putative (GT) define biosynthetic pathways for all measured glycans, chain...
The production of high-value biopharmaceuticals is dominated by mammalian cells, particularly Chinese hamster ovary (CHO) which have been widely used and preferred in manufacturing processes. discovery CRISPR-Cas9 significantly accelerated cell line engineering advances, allowing for yield quality improvements. Since then, several other CRISPR systems become appealing genome editing tools, such as the Cas12a nucleases, provide broad capabilities while utilizing short guide RNAs (gRNAs) that...
Here we develop a novel approach allowing the noncovalent assembly of proteins on well-defined nanoscaffolds such as virus particles. The antibody-binding peptide Z33 was genetically fused to monomeric yellow fluorescent protein and 4-coumarate:CoA-ligase 2. This "tag" allowed their patterning surface zucchini mosaic by means specific antibodies directed against coat virus. validated affinity assays correlative microscopy. coverage efficiency ≈ 87%. Fluorescence enzymatic activity were fully...
Abstract CRISPR-Cas12a systems are becoming an attractive genome editing tool for cell engineering due to their broader capabilities compared CRISPR-Cas9 counterparts. As opposed Cas9, the Cas12a endonucleases characterized by a lack of trans-activating crRNA (tracrRNA), which reduces complexity system and simultaneously makes CRISPR RNA (crRNA) promising approach toward further improving modulating activity systems. Here, we design validate sixteen types structurally engineered crRNAs...