Chimeric antigen receptors (CARs) are designed and engineered in a modular fashion that typically consists of an extracellular target-binding domain, a hinge region, a transmembrane domain that anchors the CAR to the cell membrane, and one or more intracellular domains that transmit activation signals. Depending on the number of costimulatory domains, Designed and engineered CARs can be classified into first (CD3ζ only), second (one costimulatory domain + CD3ζ), or third generation CARs (more than one costimulatory domain + CD3ζ).

Chimeric antigen receptor (CAR) proteins consist of an extracellular antigen-binding domain, a hinge, a transmembrane domain, a co-stimulatory domain, and an activation domain. The extracellular portion of the CAR is most often composed of a single-chain variable fragment (scFv) molecule, though camelid nanobodies and humanized natural ligands of cytokines have been used. The intracellular regions, which function in activation upon antigen binding, typically harbor a T cell activation domain derived from the CD3ζ chain of the T cell receptor. Co-stimulatory domains often include CD28 or 4-1BB and can influence CAR-T cell memory, phenotype, and metabolism. Other co-stimulatory domains have been tested including OX40, CD27, and inducible T cell co-stimulator (ICOS). The activation domain, composed of CD3ζ, DAP12, or other sequences, contained immunoreceptor tyrosine-based activation motif (ITAM) regions that can be mutated to attenuate downstream activity.
Chimeric antigen receptor (CAR) domain components.

Common co-stimulatory molecules utilized in chimeric antigen receptors (CARs).

CSMLigand(s)Biological characteristics
Expressed in resting and activated T cells. Promotes quick activation, cytokine production and slight proliferation through the PI3-kinase-AKT pathway. Promotes effector memory phenotype with high glycolytic metabolism and diminished persistence in CAR-T cells. Can promote antigen– independent exhaustion. Might favor CD4 + T cell expansion.
Involved in T cell activation and persistence by recruiting TRAF. In CAR-T cells is associated with a delayed stimulation but greater proliferative capacity compared to CD28. It is associated with a central memory phenotype and increased oxidative metabolism and persistence. Inhibits antigen - independent exhaustion in CAR-T cells.
Cooperates with 4-1BB signaling in CAR-T cell function and persistence. ICOS positioning proximal to the cell membrane can remarkably augment the effector function and persistence of CAR-T cells. It is associated with Th1 and Th17 phenotypes.
Highly expressed in activated T cells. Stimulates T cell proliferation and cytokine production. Contributes to regulatory T cells (Treg) suppression.
CD27CD70Plays a role in survival of activated CAR-T cells by upregulating Bcl-X(L) proteins. Loss of the co-stimulatory molecules CD27 and CD28 is a common phenomenon during T cell senescence.
CD40 signaling increases proliferation and secretion of cytokines in CAR-T cells. Expression of CD40 in T cells is restricted to activated T cells.
KIR2DS2Ligand uncharacterized Killer cell immunoglobulin-like receptor (KIR) KIR2DS2 induces natural killer (NK) cell activation upon ligation and is associated with protection against certain cancers and viral infections.

Oak Biosciences could construct any customized CAR via combination of your picked components.

Binding domainsHingeTransmembrane domainsCo-StimulationActivation domain
Component Candidate*scFv, VHH, VH, Cytokines, Ligands, PeptidesCD8, CD28, IgG1, IgG4CD3ζ, CD4, CD8α, CD28, ICOS4-1BB, CD27, CD28, ICOS, KIR2DS2, MYD88-CD40, OX40CD3ζ, DAP12, FcγR, ITAM mutation
2nd generation CARpick 1 or 1+Pick 1pick 1pick 1pick 1
3rd generation CARpick 1 or 1+pick 1pick 1pick 2 and 2+pick 1
*Chimeric antigen receptor (CAR) domain components could be combined to make customized 2nd or 3rd generation CAR; For next generation CAR, or dual CARs, more than one binding domains are needed.

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Customized Chimeric Antigen Receptor (CAR) Design and Construction Services

62101Second generation CAR design and construction1 ProjectContact Us
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