Identification and evaluation of novel peptide ligands specific to human CD3 epsilon
Inventors
Assignees
University of Alabama in Huntsville
Publication Number
US-12006373-B1
Publication Date
2024-06-11
Expiration Date
2041-06-16
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Abstract
The present disclosure is directed to anti-CD3ε peptides, compounds, and methods of using same. A biopanning technique was used to identify several phage clones displaying unique anti-CD3ε peptide sequences. The anti-CD3ε peptide sequences bind to human CD3ε on T cells, allowing the identification, labeling, and delivery of cargo to these T cells. Applications for the disclosed peptides include the labeling of T cells with magnetic particles for MRI detection and targeting of T cells with cargo-laden anti-CD3ε peptides for immunotherapy. Further still, incubation of T cells with anti-CD3ε peptides may result in the activation and increase production of T cells, which may in turn be harvested for applications such as CAR-T therapy.
Core Innovation
The invention provides novel anti-CD3ε peptides, compounds, and methods for their use, developed through a peptide phage display biopanning technique. Several phage clones displaying unique anti-CD3ε peptide sequences were identified, with peptides such as WSLGYTG (SEQ ID NO: 13) and FWSSPQM (SEQ ID NO: 11) demonstrating a high affinity and specificity in binding to human CD3ε on T cells. These peptides can be synthesized and functionalized with spacers and attachment groups, enabling their conjugation to various labels or cargo.
The addressed problem is the lack of small, high-specificity peptide ligands for human CD3ε—an important T cell receptor complex protein—where existing monoclonal antibodies (mAbs) suffer from large size, costly manufacturing, and limited conjugation options. The invention responds to drawbacks associated with antibodies by providing simpler, smaller peptide ligands that retain specificity to CD3ε and can be easily synthesized, modified, and conjugated.
The disclosed peptides can selectively bind CD3ε on human T cells, allowing for T cell identification, labeling, and delivery of cargo. These peptides can be conjugated to particles or molecules, such as magnetic labels or therapeutic cargo, supporting applications in cell labeling for imaging and targeted immunotherapy. Furthermore, incubation of T cells with anti-CD3ε peptides can activate T cells and enhance their proliferation, potentially supplying expanded T cell populations for further clinical use, such as CAR-T therapy.
Claims Coverage
There are four inventive features defined by the independent claims.
Anti-CD3ε peptide ligand represented by SEQ ID NO: 13
A peptide ligand specific for binding human cluster of differentiation 3 epsilon (CD3ε), wherein the ligand is defined by the amino acid sequence WSLGYTG (SEQ ID NO: 13). This inventive feature focuses on the identification and utilization of a specific peptide sequence with binding specificity to CD3ε on human T cells.
Peptide ligand with PEG6-lysine-biotin modification
A peptide ligand with binding specificity for human CD3ε, which has the structure SEQ ID NO: 31-PEG6-lysine-biotin. This inventive feature encompasses the use of the anti-CD3ε peptide in a formula where it is coupled to a polyethylene glycol spacer, a lysine residue, and a biotin group, providing options for conjugation or further modification.
Peptide ligand with maleimide-PEG2-biotin modification
A peptide ligand for binding human CD3ε, with the formula SEQ ID NO: 32-maleimide-PEG2-biotin. This inventive feature covers the anti-CD3ε peptide conjugated to a maleimide group, a PEG2 spacer, and biotin, allowing for versatile attachment to other molecules or surfaces.
Peptide ligand coupled to a labeling molecule
A peptide ligand for human CD3ε, represented by SEQ ID NO: 13, that is further coupled to a labeling molecule via an attachment group. The feature includes the optionally attached labeling molecule, which may be a fluorescent label, enzyme label, chromogenic label, luminescence label, radiation label, magnetic label, metal complex, metal, or colloidal gold, thus enabling the detection, imaging, or manipulation of T cells.
The claims collectively cover specific anti-CD3ε peptide ligands, as well as defined chemical modifications and conjugation to various labels, establishing broad and flexible means to use these peptides in binding, detecting, or manipulating human CD3ε-expressing cells.
Stated Advantages
The peptides are smaller and simpler than monoclonal antibodies, allowing for easier synthesis and modification.
The disclosed peptides can be readily conjugated to various biomaterials and labels in a defined orientation using commercially available crosslinkers.
They provide specific binding to human CD3ε, enabling targeted identification, labeling, and delivery of cargo to T cells.
Application of the peptides can activate T cells and increase T cell production without significant upregulation of activation markers or induction of activation-induced apoptosis.
Documented Applications
Labeling of T cells with magnetic particles for MRI detection.
Targeting of T cells with cargo-laden anti-CD3ε peptides for immunotherapy.
Activation and increase of T cell production for harvesting and use in applications such as CAR-T therapy.
Identification or labeling of T cells for imaging using labels such as fluorescent, enzyme, chromogenic, luminescent, radiation, magnetic, metal complexes, metal, or colloidal gold.
Production or increase of T cells via activation for downstream use, including treatment of diseases or conditions associated with T cells such as cancer (e.g., T cell lymphoma) or autoimmune disease (e.g., multiple sclerosis).
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