A5046819192- Immunotherapy and Immune Responses
- T-cell and B-cell Immunology
- Monoclonal and Polyclonal Antibodies Research
- vaccines and immunoinformatics approaches
- Immune Cell Function and Interaction
- CAR-T cell therapy research
- Peptidase Inhibition and Analysis
- Protein purification and stability
- Advanced biosensing and bioanalysis techniques
- thermodynamics and calorimetric analyses
- Synthesis and Biological Evaluation
- Computational Drug Discovery Methods
- Transgenic Plants and Applications
- Adenosine and Purinergic Signaling
- Protein Structure and Dynamics
- Antimicrobial Peptides and Activities
Constructor University
2019-2022
University of Bremen
2020
Technical University of Munich
2019
European Molecular Biology Laboratory
2019
Technical University of Denmark
2019
Disulfide-stabilized MHC class I are empty peptide-receptive molecules that rapidly load peptide and improve T cell detection. See the related Research Article by Moritz et al . in this issue.
Abstract Differential scanning fluorimetry (DSF) using the inherent fluorescence of proteins (nDSF) is a popular technique to evaluate thermal protein stability in different conditions (e.g. buffer, pH). In many cases, ligand binding increases and often this can be detected as clear shift nDSF experiments. Here, we affinity quantification based on shifts. We present four systems with ligands, ranging from nM high μM. Our study suggests that affinities determined by isothermal analysis are...
Abstract Major Histocompatibility Complex (MHC) class I molecules selectively bind peptides for presentation to cytotoxic T cells. The peptide-free state of these is not well understood. Here, we characterize a disulfide-stabilized version the human molecule HLA-A*02:01 that stable in absence peptide and can readily exchange cognate peptides. We present X-ray crystal structures HLA-A*02:01, together with have dipeptides bound A F pockets. These structural snapshots reveal amino acid side...
Functionally empty and peptide-receptive soluble HLA-A*02:01 enables high-throughput analysis of TCR binding to peptide-MHCs. See the related Research Article by Saini et al . in this issue.
Abstract An essential element of adaptive immunity is selective binding peptide antigens by major histocompatibility complex (MHC) class I proteins and their presentation to cytotoxic T lymphocytes. Using native mass spectrometry, we analyze the peptides an empty disulfide-stabilized HLA-A*02:01 molecule and, due its unique stability, determine affinities complexes loaded with truncated or charge-reduced peptides. We find that two anchor positions can be stabilized independently, further...
Abstract The peptide-dependent stability of MHC class I molecules poses a substantial challenge for their use peptide-MHC multimer-based approaches to comprehensively analyze T cell immunity. We demonstrate here the generation, analysis, and empty-loadable (peptide-free) tetramers made from disulfide-stabilized molecules. A disulfide bond links α1 α2 helices molecule at extreme end F pocket. It allows in vitro folding recombinant A2 with dipeptide subsequent removal yield stable...
Abstract Major histocompatibility complex (MHC) class I multimers have been widely used to identify antigen specific T-cells for immune monitoring, epitope discovery, and T-cell isolation. A bottleneck many peptide-MHC driven applications interrogation is the peptide ligand dependent stability of MHC proteins, which thus compels high-affinity in-vitro folding each protein use a peptide-exchange technology investigate antigens interest. To overcome this challenge, we demonstrate empty...