A5034968393- Monoclonal and Polyclonal Antibodies Research
- RNA and protein synthesis mechanisms
- Advanced Biosensing Techniques and Applications
- Glycosylation and Glycoproteins Research
- Bacterial Genetics and Biotechnology
- T-cell and B-cell Immunology
- Bacteriophages and microbial interactions
- Cytokine Signaling Pathways and Interactions
- Galectins and Cancer Biology
- interferon and immune responses
- Viral Infectious Diseases and Gene Expression in Insects
- Autoimmune Bullous Skin Diseases
- Antimicrobial Resistance in Staphylococcus
- Influenza Virus Research Studies
- CAR-T cell therapy research
- Transgenic Plants and Applications
- Plant biochemistry and biosynthesis
- Advanced biosensing and bioanalysis techniques
- Enzyme Structure and Function
- Immune Response and Inflammation
- Toxin Mechanisms and Immunotoxins
- Immune Cell Function and Interaction
Czech Academy of Sciences, Institute of Biotechnology
2011-2024
Institut de Biologie Structurale
2017
Czech Academy of Sciences
2015
Recombinant ligands derived from small protein scaffolds show promise as robust research and diagnostic reagents next generation therapeutics. Here, we high-affinity binders of human interferon gamma (hIFNγ) the three helix bundle scaffold albumin-binding domain (ABD) G Streptococcus G148. Computational interaction energy mapping, solvent accessibility assessment, in silico alanine scanning identified 11 residues surface ABD suitable for randomization. A corresponding combinatorial library...
Interferon-γ receptor 2 is a cell-surface that required for interferon-γ signalling and therefore plays critical immunoregulatory role in innate adaptive immunity against viral also bacterial protozoal infections. A crystal structure of the extracellular part human (IFNγR2) was solved by molecular replacement at 1.8 Å resolution. Similar to other class receptors, IFNγR2 has two fibronectin type III domains. The characteristic structural features are concentrated its N-terminal domain: an...
Engineered small non-antibody protein scaffolds are a promising alternative to antibodies and especially attractive for use in therapeutics diagnostics. The advantages include smaller size more robust, single-domain structural framework with defined binding surface amenable mutation. This calls systematic approach designing new suitable one or methods of directed evolution. We hereby describe process based on an analysis structures from the Protein Data Bank their experimental examination....
We describe a computer-based protocol to design protein mutations increasing binding affinity between ligand and its receptor. The method was applied mutate interferon-<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M3"><mml:mrow><mml:mi>γ</mml:mi></mml:mrow></mml:math>receptor 1 (IFN-<mml:math id="M4"><mml:mrow><mml:mi>γ</mml:mi></mml:mrow></mml:math>-Rx) increase natural IFN-<mml:math id="M5"><mml:mrow><mml:mi>γ</mml:mi></mml:mrow></mml:math>, important for innate immunity....
We present a novel surface plasmon resonance (SPR) biosensor for detection of human interferon gamma (hIFNγ), an important diagnostic marker several infectious diseases. The employs engineered proteins derived from albumin binding domain (ABD) and selected with ribosome display to provide high affinity towards hIFNγ. demonstrate that the provides rapid, sensitive, cost-effective platform hIFNγ allows its at nanomolar levels both in buffer diluted plasma.
Combining computational and experimental tools, we present a new strategy for designing high affinity variants of binding protein. The is increased by mutating residues not at the interface, but positions lining internal cavities one interacting molecules. Filling lowers flexibility protein, possibly reducing entropic penalty binding. approach was tested using interferon-γ receptor 1 (IFNγR1) complex with IFNγ as model. Mutations were selected from 52 amino acid IFNγR1 protocol based on...
We combined cell-free ribosome display and cell-based yeast selection to build specific protein binders the extracellular domain of human interleukin 9 receptor alpha (IL-9Rα). The target, IL-9Rα, is involved in signalling pathway IL-9, a pro-inflammatory cytokine medically important for its involvement respiratory diseases. successive use modified protocols displays allowed us combine their strengths-the virtually infinite power production (mostly) properly folded soluble proteins display....
We hereby describe the process of design and selection nonantibody protein binders mimicking cytokine signaling. chose to mimic signaling IFN-λ1, type 3 interferon (also known as IL-29) for its novelty importance biological functions. All four interferons λ signal through binding extracellular domains IL-28 receptor 1 (IL-28R1) IL-10 2 (IL-10R2). Our were therefore trained bind both receptors simultaneously. The bifunctional binder molecules developed by yeast display, a method directed...
Here, we present a previously undescribed approach to modify N-terminal sequences of recombinant proteins increase their production yield in Escherichia coli. Prior research has demonstrated that the nucleotides immediately following start codon can significantly influence protein expression. However, impact these is construct-specific and not universally applicable all proteins. Most previous been limited selecting from few rationally designed sequences. In contrast, used directed...
Binder H33 is a small protein binder engineered by ribosome display to bind human interleukin 10. Crystals of severe diffraction anisotropy. A set data files with correction for anisotropy based on different local signal-to-noise ratios was prepared. Paired refinement used find the optimal anisotropic high-resolution limit data: 3.13–2.47 Å. The structure belongs 2% crystal structures highest solvent content in Protein Data Bank.