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This pair of synthetic peptides spontaneously self-assembles into a biomaterial with a precise supramolecular architecture with functional capabilities. The biomaterials market is expected to reach $150 billion by 2021. However, methods of producing advanced biomaterials with immobilized folded proteins remain a challenge, which limits opportunities to create biomaterials with advanced functional properties like catalysis, antigen conformation or molecular recognition.
Researchers at the University of Florida have developed a pair of oppositely charged synthetic peptides, called CATCH, or Co-Assembly Tags based on Charge-complementarity, to immobilize folded proteins into supramolecular biomaterials. Either of the CATCH peptides attaches to a protein of interest as a recombinant fusion tag. Proteins harboring a CATCH tag co-assemble with CATCH peptides into nanofibers, resulting in supramolecular biomaterials with a non-covalently immobilized protein displayed on their surface. The versatility of using a recombinant fusion tag for protein immobilization makes this platform potentially useful for various biomedical and biotechnological applications, including tissue engineering, regenerative medicine, drug delivery, immunotherapy, immunomodulation, and biosensing.
A synthetic peptide pair that facilitates non-covalent protein immobilization into biomaterials for applications ranging from drug delivery to tissue engineering
The CATCH peptides that form the basis for the supramolecular hydrogel can be synthesized and purified in high yield using both conventional solid-phase and recombinant expression methods. The latter provides a notable advantage over synthetic biomolecules capable of self-assembly. Because CATCH peptides co-assemble into ß-sheet nanofibers only when combined, due to electrostatic interactions, CATCH fusion proteins can be expressed and recovered from the soluble phase in exceptionally high yield and purity when compared to other platforms. CATCH peptides and fusion proteins can also be used to fabricate micron-sized particles or macroscopic hydrated gels by simply varying the composition of peptides and fusion proteins present during assembly.
This fusion protein is composed of galectin-1 and galectin-3 and is a candidate therapeutic for modulating immune responses and treating inflammation. The global autoimmune disease therapeutics market should reach $1.5 billion by 2025. Galectins are promising therapeutic candidates for managing immune responses and inflammation because they can alter the phenotype and function of immune cells. Currently, galectin-1 and galectin-3 variants are used separately as therapeutics, but both require high doses that are expensive and impractical for clinical use.
Researchers at the University of Florida have developed a fusion protein composed of galectin-1 and galectin-3 that is a candidate therapeutic for regulating immune responses and controlling inflammation. This new candidate therapeutic displays greater binding ability, better immunomodulatory potency, and requires lower doses to be effective than other galectin therapeutics.
New galectin therapeutic with lower minimum effective dose and better binding in immune response regulation than other galectin therapeutics
This candidate therapeutic for regulating immune responses and controlling inflammation is a fusion protein comprised of one galectin-1 polypeptide and one galectin-3 polypeptide. This protein dimer has a better binding affinity and lower effective dosing requirement than current galectin candidate therapeutics, and it was more effective than other galectin therapeutics at inducing T-cell apoptosis critical for regulating immune responses.
This bifunctional fusion protein resets the immune system to promote healthy tissue function by anchoring enzymes to the site of inflammation. Biologic drugs used to treat inflammatory diseases are a market that exceeds $30 billion annually. Available treatments to suppress inflammation are systemic, affecting unintended tissue and causing significant side effects. Localizing immunosuppressive drugs to sites of inflammation eliminates those problems.
Researchers at the University of Florida have developed a new approach to localize immunosuppressive enzymes to sites of inflammation. This technology platform is known as Galectin Anchors for Therapeutic Enzyme Retention. Specifically, they create a bifunctional fusion of an enzyme and galectin-3, a protein that binds to sugars decorating every tissue in our bodies. By binding to tissue sugars, Galectin-3 anchors the enzyme at the injection site. This prevents enzyme diffusion into surrounding tissue or entry into circulation. Their data show anchored enzyme formulations that persist at sites of inflammation for up to 2 weeks. Anchoring enzymes at the site of inflammation eliminates the side-effects that result from systemic distribution of drugs throughout the entire body.
The immunosuppressive enzyme utilized is indoleamine 2,3-dioxygenase (IDO), which breaks down the essential amino acid tryptophan into kynurenines. IDO naturally regulates the immune system in various contexts. For example, IDO establishes local maternal tolerance to the fetus to protect from immune attack without increasing host vulnerability to infection. Our bifunctional fusion protein resets the immune system to promote healthy tissue function by anchoring IDO to the site of inflammation, where it is administered. Restricting distribution of IDO avoids off-target side-effects and systemic immunosuppression.
Bifunctional fusion protein of galectin-3 and indoleamine 2,3-dioxygenase locally suppress inflammation
The bifunctional fusion protein, Gal3-IDO binds to tissue glycans to anchor the immunosuppressive enzyme at sites of inflammation. The inventors have demonstrated in in vivo models of osteoarthritis that Gal3-IDO can reduce inflammation, relieve pain, and restore gait. The inventors have demonstrated in in vivo models of periodontal disease that Gal3-IDO- can reduce inflammation and prevent the bone loss that leads to the need for tooth extraction. The inventors have demonstrated in in vivo models of bacterial endotoxin challenge that Gal3-IDO can shut down inflammation locally without systemic suppression.
This polymer crosslinked, more stable Galectin-1 homodimer suppresses inflammation more potently than unmodified recombinant Galectin-1. Galectin-1 (Gal1) modulates innate and adaptive immunity by regulating dendritic cell migration, inducing activated T cell apoptosis, and biasing Th1 and Th17 T-cell subsets toward immunosuppressive phenotypes. Prior work has shown that recombinant Gal1 suppresses T-cell-dependent chronic inflammation in diseases such as arthritis, hepatitis, and colitis. To retain its immune-modulating properties, Gal1 must adopt a homodimer conformation, which is an unstable structure. Using the synthetic polymer poly(ethylene glycol) (PEG) as a crosslinker provides a stable Gal1 homodimer. PEG is advantageous over other chemistries because it is well-tolerated by humans and has a proven track record as a therapeutic protein appendage.
A stable, PEG-crosslinked Gal1 homodimer as an immunomodulatory therapeutic agent for chronic inflammatory and autoimmune diseases
Translation of Galectin-1 (Gal1) therapies from bench to bedside has been unsuccessful. Gal1 administration is difficult because it typically requires high doses that favor the formation of the active homodimer conformation. This technology creates a more stable and potent therapeutic Gal1 by employing the synthetic polymer PEG as a crosslinker to stabilize homodimer formation. Although an abundance of pegylation exists in the protein-engineering field, the novelty of this system is using PEG with acrylates or maleimides at either end of polymer to attach onto two separate Gal1 monomers to create a stabilized dimeric structure.
