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Anti-CD33 (C-20) h(4-1BB-CD3ζ) CAR-T Cells [Mouse CAR-T Cells] (MCAR-P1158)

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All products and services are For Research Use Only and CANNOT be used in the treatment or diagnosis of disease.

Adoptive immunotherapy using T cells expressing chimeric antigen receptors (CARs) has produced remarkable clinical outcomes. However, much of the mechanisms of action, such as the development of memory responses and sources of immune cytokines, remain elusive largely due to the challenge of characterizing human CAR T cell function in vivo. Creative Biolabs has established a syngeneic mouse model with a functional mouse immune system and genetically-matched (autologous) Acute lymphoblastic leukemia (ALL) that permits modeling of CD33-targeted CAR T cell therapy in immunocompetent hosts without allogeneic immune responses. Our anti-CD33 CAR T cells are prepared from freshly isolated T lymphocytes in a syngeneic mouse model, followed by ex vivo transduction with anti-CD33 CAR lentiviral particles and amplification process to generate CAR T cells. These syngeneic mouse model-derived CAR-T cells offer many advantages including permitting the evaluation of antitumor responses in immunocompetent murine hosts, testing human CAR-T constructs against human primary malignance, and preclinical evaluation of CAR T cell therapies.

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T Cell Background

  • Target
  • CD33
  • Applications
  • The mouse CD33 CAR-T cells can be used for evaluation of the safety, toxicity, persistence and exhaustion of CAR T cells in syngeneic mouse model.
  • T Cell Source
  • Syngeneic Mouse Model
  • Donor Mouse Strain
  • BALB/c
  • Donor Tumor Cell Line
  • Acute lymphoblastic leukemia (ALL) Cell Line
  • Mouse Modeling
  • Syngeneic mouse models can be a powerful tool for testing immunotherapies. They consist of tumor tissues from the same genetic background as the given immuno-competent mouse strain. A syngeneic mouse model provides an effective approach for studying how cancer therapies perform in the presence of a functional immune system.

    At Creative Biolabs, we have established syngeneic models by injecting a recipient of a specific genetic background with cell lines previously established through isolation of tumor cells from a mouse of the same genetic background. Our syngeneic models that have been fully characterized with known checkpoint inhibitors (e.g., anti-PDL-1, anti-PD-1, anti-CTLA-4), whole exome sequencing, and immunologic profiling data, backed by experienced scientists who can ensure the model runs smoothly.
  • T Cell Isolation
  • The mouse T cells were isolated from syngeneic mouse spleens by MACS using anti-mouse CD3 micro-beads.

CAR Design

  • CAR Introduction
  • The vector of anti-CD33 chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target Mouse CD33. The T cells are genetically modified through transduction with a lentiviral vector expressing scFv of anti-CD33 antibody linked to 4-1BB and CD3ζ signaling domains. And the vector product was designed for the treatment of Acute lymphoblastic leukemia (ALL).
  • scFv Clone
  • C-20
  • Target Species
  • Mouse
  • scFv Host Species
  • Humanized
  • CAR Construction
  • scFv-4-1BB-CD3ζ
  • CAR Signaling Cassetes
  • 41BB
    CD137 (also known as 4-1BB) is a surface co-stimulatory glycoprotein originally described as present on activated T lymphocytes, which belongs to the tumor necrosis factor (TNF) receptor superfamily. It is expressed mainly on activated CD4+ and CD8+ T cells, and binds to a high-affinity ligand (4-1BBL) expressed on several antigen-presenting cells such as macrophages and activated B cells. On the basis of preclinical observation, this molecule can promote the persistence of antigen-specific and antigennonspecific chimeric antigen receptor T-cells to significantly increases antitumor activity.
    CD3ζ
    CD3ζ, also known as T-cell receptor zeta, which together with T-cell receptor and CD3γ, δ , ε chain, forms the TCR-CD3 complex. ζ was expressed independently from the complex. The zeta chain plays an important role in coupling antigen recognition to several intracellular signal-transduction pathways. CD3-zeta, which contains 3 ITAMs, is the most commonly used endodomain component of CARs. It transmits an activation signal to the T cell after antigen is bound. CD3-zeta may not provide a fully competent activation signal and additional co-stimulatory signaling is needed. For example, chimeric 4-1BB and OX40 can be used with CD3-zeta to transmit a proliferative/survival signal, or all three can be used together.
  • CAR Vector Name
  • pCDCAR1
  • CAR Vector Length
  • ~8kb
  • CAR Vector Type
  • Lentiviral vector
  • CAR Generation
  • Second
  • Targeting Diseases
  • Acute lymphoblastic leukemia (ALL)

CAR-T Cell

  • CAR-T Cell Preparation
  • Freshly mouse T cells were isolated from mouse spleens by MACS using anti-mouse CD3 micro-beads, and then transduced with anti-CD33 CAR-expressing lentiviral particles.
  • CAR-T Cell Expansion
  • The CAR virus transduced T cells were then expanded for about 2 weeks using our in-house protocol. The expanded CAR-T cells should be used immediately in mice.
  • CAR-T Cell Characterization
  • The expression of mouse CD33 is characterized by flow cytometry.

Quality Control

  • Cell Purity
  • >95%
  • Cell Viability
  • >90%
  • Mycoplasma Testing
  • The cell line has been screened using the luciferase based mycoplasma detection kit to confirm the absence of mycoplasma species.
  • Sterility Testing
  • Creative Biolabs provides sterility testing in accordance with USP and EP regulations. All of our sterility testing is performed in an isolator or clean room environments. The cell line has been screened using the membrane filtration testing methods to confirm the absence of aerobic, anaerobic and fungi microorganisms.

Shipping and Handling

  • Shipping
  • Dry Ice
  • Storage
  • Lyophilized cells should be stored in a liquid nitrogen tank (-150°C~-190°C). Once reconstituted, the cells may be used for up to five days if properly stored at 2°C - 20°C in the buffer provided.
  • Handling Notes
  • Frozen cells should be thawed immediately upon receipt and grown according to handling procedure to ensure cell viability and proper assay performance.
    Note: Do not freeze the cells upon receipt as it may result in irreversible damage to the cell line.
    Disclaimer: We cannot guarantee cell viability if the cells are not thawed immediately upon receipt and grown according to handling procedure.
  • Warnings
  • Avoid multiple freeze/thaw cycles
  • Research Use Only
  • For research use only, not for diagnostic or therapeutic use.

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For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.

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