A5022774854- Monoclonal and Polyclonal Antibodies Research
- Glycosylation and Glycoproteins Research
- Advanced Proteomics Techniques and Applications
- Advanced Biosensing Techniques and Applications
- Ubiquitin and proteasome pathways
- Cancer, Lipids, and Metabolism
- Peptidase Inhibition and Analysis
- SARS-CoV-2 and COVID-19 Research
- Biochemical and Structural Characterization
- Protein Degradation and Inhibitors
- Immunotherapy and Immune Responses
- Hepatitis C virus research
- COVID-19 Clinical Research Studies
- Chemical Synthesis and Analysis
- Ferroptosis and cancer prognosis
- Autophagy in Disease and Therapy
- Fibroblast Growth Factor Research
- Long-Term Effects of COVID-19
- RNA and protein synthesis mechanisms
- CAR-T cell therapy research
- vaccines and immunoinformatics approaches
- CRISPR and Genetic Engineering
- Mass Spectrometry Techniques and Applications
- Protein purification and stability
- Biosimilars and Bioanalytical Methods
Louisiana State University
2023-2024
University of Toronto
2017-2024
University of Oxford
2013-2020
University of Nova Gorica
2019
Université Paris Sciences et Lettres
2017
Institut Curie
2017
Venezia Tecnologie (Italy)
2010
Programmed connection of amino acids or nucleotides into chains introduced a revolution in control biological function. Reacting proteins together is more complex because the number reactive groups and delicate stability. Here we achieved sequence-programmed irreversible protein units, forming polyprotein teams by sequential amidation transamidation. SpyTag peptide engineered to spontaneously form an isopeptide bond with SpyCatcher protein. By engineering adhesin RrgA from Streptococcus...
Significance Building proteins into assemblies faces challenges in specificity and stability of the connections. Proteins are ideally connected via peptide tags for minimal disruption function. Streptococcus pyogenes contains a protein that locks itself together. After genetic dissection, we created (SpyLigase) two With on opposite ends an antibody or affibody (an antibody-like scaffold), SpyLigase assembled polyantibody polyaffibody chains. Magnetic beads can isolate specific cell types,...
The capture of biotin by streptavidin is an inspiration for supramolecular chemistry and a central tool biological nanotechnology, because the rapid exceptionally stable interaction. However, there no robust orthogonal interaction to this hub, limiting size complexity molecular assemblies that can be created. Here we combined traptavidin (a variant maximizing binding strength) with irreversible SpyTag peptide engineered form spontaneous isopeptide bond its protein partner SpyCatcher. or...
Abstract Vaccines have demonstrated remarkable effectiveness in protecting against COVID‐19; however, concerns regarding vaccine‐associated enhanced respiratory diseases (VAERD) following breakthrough infections emerged. Spike protein subunit vaccines for SARS‐CoV‐2 induce VAERD hamsters, where aluminum adjuvants promote a Th2‐biased immune response, leading to increased type 2 pulmonary inflammation animals with infections. To gain deeper understanding of the potential risks and underlying...
A comprehensive analysis of the phosphoproteome is essential for understanding molecular mechanisms human diseases. However, current tools used to enrich phosphotyrosine (pTyr) are limited in their applicability and scope. Here, we engineered new superbinder Src-Homology 2 (SH2) domains that diverse sets pTyr-peptides. We phage display select a Fes-SH2 domain variant (superFes; sFes1) with high affinity pTyr solved its structure bound pTyr-peptide. performed systematic structure–function...
Ubiquitin interacting motifs (UIMs) are short α-helices found in a number of eukaryotic proteins. UIMs interact weakly but specifically with ubiquitin conjugated to other proteins, and so doing, mediate specific cellular signals. Here we used phage display generate variants (UbVs) targeting the N-terminal UIM yeast Vps27 protein. Selections yielded UbV.v27.1, which recognized cognate high specificity relative bound an affinity more than two orders magnitude higher that ubiquitin. Structural...
Suppressor of cytokine signaling (SOCS)2 protein is a key negative regulator the growth hormone (GH) and Janus kinase (JAK)-Signal Transducers Activators Transcription (STAT) cascade. The central SOCS2-Src homology 2 (SH2) domain characteristic SOCS family proteins an important module that facilitates recognition targets bearing phosphorylated tyrosine (pTyr) residues. Here we identify exosite on SOCS2-SH2 which, when bound to non-phosphorylated peptide (F3), enhances SH2 affinity for...
Despite the revolutionary achievements of chimeric antigen receptor (CAR) T cell therapy in treating cancers, especially leukemia, several key challenges still limit its therapeutic efficacy. Of particular relevance is relapse cancer large part, as a result exhaustion and short persistence CAR-T cells vivo. IL-2-inducible kinase (ITK) critical modulator strength T-cell (TCR) signaling, while role CAR signaling unknown. By electroporation clustered regularly interspaced palindromic repeats...
Abstract Histone methylation is an important post-translational modification that plays a crucial role in regulating cellular functions, and its dysregulation implicated cancer developmental defects. Therefore, systematic characterization of histone necessary to elucidate complex biological processes, identify biomarkers, ultimately, enable drug discovery. Studying relies on the use antibodies, but these suffer from lot-to-lot variation, are costly, cannot be used live cells....
Cell isolation via antibody-targeted magnetic beads is a powerful tool for research and clinical applications, most recently isolating circulating tumor cells (CTC). Nonetheless fundamental features of the cell-bead interface are still unknown. Here we apply clinically relevant antibody against cancer target HER2 (ErbB2) cell isolation. We investigate how many proteins per sufficient to be isolated. To understand importance primary affinity, compared series point mutants with known...
Abstract Understanding protein folding under conditions similar to those found in vivo remains challenging. Folding occurs mainly vectorially as a polypeptide emerges from the ribosome or membrane translocon. Protein during translocation is particularly difficult study. Here, we describe single-molecule method characterize folded state of individual proteins after translocation, by monitoring ionic current passing through pore. We tag both N and C termini model protein, thioredoxin, with...
Ubiquitin (Ub)-binding domains embedded in intracellular proteins act as readers of the complex Ub code and contribute to regulation numerous eukaryotic processes. Ub-interacting motifs (UIMs) are short α-helical modular recognition elements whose role controlling proteostasis signal transduction has been poorly investigated. Moreover, impaired or aberrant activity UIM-containing implicated diseases, but targeting remains a major challenge. To overcome this limitation, we developed variants...
Abstract USP37 is a deubiquitinase (DUB) with roles in the regulation of DNA damage repair and cohesion sister chromatids during mitosis. contains unique insert three ubiquitin interacting motifs (UIMs) within its catalytic DUB domain. We investigated role UIMs ability to cleave di-ubiquitin chains. found that third UIM recognizes proximal moiety K48 di-Ub potentiate cleavage activity posit this mechanism action may be generalizable other chain types. In case K48-linked chains potentiation...
Background: Adhirons are small (10 kDa) synthetic ligands that might represent an alternative to antibody fragments and scaffolds such as DARPins or affibodies. Methods: We prepared a conceptionally new adhiron phage display library allows the presence of cysteines in hypervariable loops successfully panned it against antigens possessing different characteristics. Results: recovered binders specific for membrane epitopes plant cells by panning directly pea protoplasts soluble C-Reactive...
Histidine phosphorylation (pHis) is a non-canonical post-translational modification (PTM) that historically understudied due to lack of robust reagents are required for its investigation, such as high affinity pHis-specific antibodies. Engineering antibodies very challenging the labile nature phosphoramidate (P-N) bond and stringent requirements selective recognition two isoforms, 1-phosphohistidine (1-pHis) 3-phosphohistidine (3-pHis). Here, we present strategy