A5077717500- Monoclonal and Polyclonal Antibodies Research
- HER2/EGFR in Cancer Research
- Radiopharmaceutical Chemistry and Applications
- CAR-T cell therapy research
- Glycosylation and Glycoproteins Research
- Protease and Inhibitor Mechanisms
- Biochemical and Structural Characterization
- Cancer Immunotherapy and Biomarkers
- Blood properties and coagulation
- Viral Infectious Diseases and Gene Expression in Insects
- Venomous Animal Envenomation and Studies
- Enzyme Structure and Function
- Virus-based gene therapy research
- Erythrocyte Function and Pathophysiology
- Signaling Pathways in Disease
- Nanofabrication and Lithography Techniques
- Immunotherapy and Immune Responses
- Cellular transport and secretion
- Immune Cell Function and Interaction
- Complement system in diseases
- T-cell and B-cell Immunology
- Blood Coagulation and Thrombosis Mechanisms
- Digital Storytelling and Education
- Protein Interaction Studies and Fluorescence Analysis
- Cancer-related gene regulation
Aarhus University
2015-2024
Novo Nordisk (Denmark)
2020-2021
The costimulation of immune cells using first-generation anti-4-1BB monoclonal antibodies (mAbs) has demonstrated anti-tumor activity in human trials. Further clinical development, however, is restricted by significant off-tumor toxicities associated with FcγR interactions. Here, we have designed an Fc-free tumor-targeted 4-1BB-agonistic trimerbody, 1D8N/CEGa1, consisting three single-chain variable fragments and anti-EGFR single-domain positioned extended hexagonal conformation around the...
The redirection of T cell activity using bispecific antibodies is one the most promising cancer immunotherapy approaches currently in development, but it limited by cytokine storm-related toxicities, as well pharmacokinetics and tumor-penetrating capabilities current antibody formats. Here, we have engineered ATTACK (Asymmetric Tandem Trimerbody for Activation Cancer Killing), a novel cell-recruiting which combines three EGFR-binding single-domain (VHH; clone EgA1) with single CD3-binding...
Retargeting of T lymphocytes toward cancer cells by bispecific antibodies has demonstrated its therapeutic potential, with one such antibody approved for the treatment acute lymphoblastic leukemia (blinatumomab) and several other in clinical trials. However, improvement their efficacy selectivity solid tumors is still required. Here, we describe a novel tandem T-cell recruiting trispecific colorectal (CRC). This construct, termed engager (TriTE), consists CD3-specific single-chain Fv (scFv)...
Chimeric antigen receptor (CAR)-modified T cells have revolutionized the treatment of CD19-positive hematologic malignancies. Although anti-CD19 CAR-engineered autologous can induce remission in patients with B-cell acute lymphoblastic leukemia, a large subset relapse, most them disease. Therefore, new therapeutic strategies are clearly needed. Here, we report comprehensive study comparing engineered either expressing second-generation CAR (CAR-T19) or secreting CD19/CD3-targeting bispecific...
Immune checkpoint blockade (ICB) with antibodies has shown durable clinical responses in a wide range of cancer types, but the overall response rate is still limited. Other effective therapeutic modalities to increase ICB rates are urgently needed. New bispecific antibody (bsAb) formats combining effect and direct action on cells could improve efficacy current immunotherapies. Here, we report development PD-L1/EGFR symmetric bsAb by fusing dual-targeting tandem trimmer body human IgG1 hinge...
Abstract Here, we describe a new strategy that allows the rapid and efficient engineering of mono multispecific trivalent antibodies. By fusing single-domain antibodies from camelid heavy-chain-only immunoglobulins (V HHs ) to N-terminus human collagen XVIII trimerization domain (TIE produced monospecific trimerbodies were efficiently secreted as soluble functional proteins by mammalian cells. The purified V HH -TIE trimeric in solution exhibited excellent antigen binding capacity....
Human α2-macroglobulin (A2M) is the most characterized protease inhibitor in alpha-macroglobulin (αM) superfamily, but structure of its native conformation has not been determined. Here, we combined negative stain electron microscopy (EM), small-angle X-ray scattering (SAXS), and cross-linking-mass spectrometry (XL-MS) to investigate A2M collapsed conformations that are obtained through aminolysis thiol ester by methylamine or cleavage bait region trypsin. The interpretation these data...
Abstract Purpose: The induction of 4-1BB signaling by agonistic antibodies can drive the activation and proliferation effector T cells thereby enhance a T-cell–mediated antitumor response. Systemic administration anti-4-1BB–agonistic IgGs, although effective preclinically, has not advanced in clinical development due to their severe hepatotoxicity. Experimental Design: Here, we generated humanized EGFR-specific 4-1BB-agonistic trimerbody, which replaces IgG Fc region with human collagen...
Costimulation of tumor-infiltrating T lymphocytes by anti-4-1BB monoclonal antibodies (mAbs) has shown anti-tumor activity in human trials, but can be associated with significant off-tumor toxicities involving FcγR interactions. Here, we introduce albumin-fused mouse and bispecific clinically favorable pharmacokinetics designed to confine 4-1BB costimulation the tumor microenvironment. These Fc-free agonists consist an EGFR-specific VHH antibody, a 4-1BB-specific scFv, albumin sequence...
4-1BB (CD137) is an inducible costimulatory receptor that promotes expansion and survival of activated T cells; IgG-based 4-1BB-agonistic monoclonal antibodies exhibited potent antitumor activity in clinical trials. However, the development those restricted by major off-tumor toxicities associated with FcγR interactions. We have recently generated EGFR-targeted trimerbody demonstrated strong did not induce systemic inflammatory cytokine secretion hepatotoxicity first-generation agonists....
Adoptive transfer of genetically engineered human cells secreting bispecific T-cell engagers has shown encouraging therapeutic effects in preclinical models cancer. However, reducing the toxicity and improving effectiveness this emerging immunotherapeutic strategy will be critical to its successful application. We have demonstrated that for gene-based antibody strategies, two-chain diabodies a better safety profile than single-chain tandem scFvs (single-chain variable fragments), because...
Human α2-macroglobulin (A2M) is an abundant protease inhibitor in plasma, which regulates many proteolytic processes and involved innate immunity. A2M's unique protease-trapping mechanism of inhibition initiated when a cleaves within the exposed highly susceptible "bait region." As wild-type bait region permissive to cleavage by most human proteases, A2M accordingly broad-spectrum inhibitor. In this study, we extensively modified order identify any potential functionally important elements...
Abstract A2ML1 is a monomeric protease inhibitor belonging to the A2M superfamily of inhibitors and complement factors. Here, we investigate protease-inhibitory mechanism human determine structures its native protease-cleaved conformations. The functional inhibitory unit monomer that depends on covalent binding (mediated by A2ML1’s thioester) achieve inhibition. In contrast tetramer which traps proteases in two internal chambers formed four subunits, disordered regions surround trapped may...
The glycosylphosphatidylinositol (GPI)-anchored protein cluster of differentiation 109 (CD109) is expressed on many human cell types and modulates the transforming growth factor β (TGF-β) signaling network. CD109 belongs to alpha-macroglobulin family proteins, known for their protease-triggered conformational changes. However, effect proteolysis its conformation are unknown. Here, we investigated interactions with proteases. We found that a diverse selection proteases cleaved peptide bonds...
Most proteins in the α-macroglobulin (αM) superfamily contain reactive thiol esters that are required for their biological function. Here, we have characterized human α2-macroglobulin (A2M) and complement component C3 mutants A2M Q975C Q1013C, which replace CGEQ ester motifs of original with disulfide-forming sequence CGEC. Mass spectrometry showed intended disulfide was formed both proteins. The correct folding native conformation were shown by its assembly to a tetramer, an initially slow...
Agonistic monoclonal antibodies (mAbs) targeting the co-stimulatory receptor 4-1BB are among most effective immunotherapeutic agents across pre-clinical cancer models. However, clinical development of full-length agonistic mAbs, has been hampered by dose-limiting liver toxicity. We have previously developed an EGFR-targeted 4-1BB-agonistic trimerbody (1D8N/CEGa1) that induces potent anti-tumor immunity without systemic toxicity, in immunocompetent mice bearing murine colorectal carcinoma...
Proteins in the α-macroglobulin (αM) superfamily use thiol esters to form covalent conjugation products upon their proteolytic activation. αM protease inhibitors theirs conjugate proteases and preferentially react with primary amines (
Protein aggregation in the outermost layers of cornea, which can lead to cloudy vision and severe cases blindness, is linked mutations extracellular matrix protein transforming growth factor-β-induced (TGFBIp). Among most frequent pathogenic are R124H R555W, both associated with granular corneal dystrophy (GCD) characterized by early-onset formation amorphous aggregates. The molecular mechanisms GCD largely unknown. In this study, we determined crystal structures R124H, lattice...
Abstract Administration of neutralizing antibodies (nAbs) has proved to be effective by providing immediate protection against SARS‐CoV‐2. However, dual strategies combining virus neutralization and immune response stimulation enhance specific cytotoxic T cell responses, such as dendritic (DC) cross‐priming, represent a promising field but have not yet been explored. Here, broadly nAb, TN , are first generated grafting an anti‐RBD biparatopic tandem nanobody onto trimerbody scaffold. Cryo‐EM...
Abstract The C3 protein is the central molecule within complement system and undergoes pattern-recognition-dependent proteolytic activation to C3b in presence of pathogens damage-associated patterns. Spontaneous pattern-independent occurs via hydrolysis, resulting C3(H 2 O). However, structural details hydrolysis remain elusive. Here, we show that conformation O) analog, C3MA, which thioester broken by aminolysis indistinguishable from except for 77-residue anaphylatoxin (ANA) domain. In...