CD40 Human, HEK
Description
Production and Quality Control
Recombinant CD40 is produced via stable transfection of HEK293 cells, followed by affinity chromatography. Critical quality metrics include:
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Bioactivity: Validated via NF-κB reporter assays (EC₅₀: 0.5–2 nM) .
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Stability: Maintains activity for 20 passages in cell lines or 2–4 weeks at 4°C .
Table 2: Functional Assays for CD40 Human, HEK
Immunotherapy Development
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CEA-CD40 bispecific antibodies: Enhance dendritic cell (DC) activation and tumor antigen cross-presentation by colocalizing CD40 agonism with carcinoembryonic antigen (CEA)+ tumors .
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EV-mediated antigen delivery: CD40-targeted extracellular vesicles (EVs) loaded with tumor antigens improve DC priming of CD8+ T cells .
Mechanistic Studies
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Oligomerization: CD40 self-assembles into dimers via CRD1, which suppresses basal signaling until ligand engagement .
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Pathway analysis: CD40 ligation activates TRAF6 and MAP3K8, driving ERK phosphorylation in B cells and macrophages .
Key Considerations for Use
Product Specs
CD40 is a member of the TNF receptor family and plays a crucial role in various immune and inflammatory responses. These include T cell-dependent immunoglobulin class switching, memory B cell development, and the formation of germinal centers. The transcription factor AKNA, which belongs to the AT-hook family, is known to regulate the expression of CD40 and its ligand, which is important for interactions between similar cells. The adaptor protein TNFR2 interacts with CD40 and acts as a signal transduction mediator. The interaction between CD40 and its ligand is believed to be essential for the activation of microglia induced by amyloid-beta, suggesting its involvement in the early stages of Alzheimer's disease pathogenesis.
Recombinant Human CD40, produced in HEK cells, is a single, glycosylated polypeptide chain consisting of 412 amino acids (specifically, amino acids 21-193). It has a molecular weight of 46.1 kDa. This CD40 protein is expressed with a 239 amino acid hIgG-His-Tag at its C-terminus and purified using proprietary chromatographic techniques.
The product is a clear, colorless solution that has been sterilized by filtration.
The CD40 protein solution has a concentration of 1 mg/ml. It is formulated in a solution containing 10% glycerol and Phosphate-Buffered Saline (PBS) at a pH of 7.4.
For short-term storage (up to 2-4 weeks), the product should be stored at 4°C. For longer storage, it is recommended to freeze the product at -20°C. For extended storage periods, adding a carrier protein (such as HSA or BSA) at a concentration of 0.1% is advisable. Repeated freezing and thawing of the product should be avoided.
The purity of the CD40 protein is greater than 95%, as determined by SDS-PAGE analysis.
CD40 Molecule, TNF Receptor Superfamily Member 5, TNFRSF5, Tumor Necrosis Factor Receptor Superfamily, Member 5, Bp50, B-Cell Surface Antigen CD40, CD40L Receptor, CDW40, B Cell Surface Antigen CD40, B Cell-Associated Molecule, CD40 Antigen (TNF Receptor Superfamily Member 5), CD40 Type II Isoform, Nerve Growth Factor Receptor-Related B-Lymphocyte Activation Molecule, p50, Tumor Necrosis Factor Receptor Superfamily Member 5, CDw40, CD40 Antigen
HEK 293.
EPPTACREKQ YLINSQCCSL CQPGQKLVSD CTEFTETECL PCGESEFLDT WNRETHCHQH KYCDPNLGLR VQQKGTSETD TICTCEEGWH CTSEACESCV LHRSCSPGFG VKQIATGVSD TICEPCPVGF FSNVSSAFEK CHPWTSCETK DLVVQQAGTN KTDVVCGPQD RLRLEPKSCD KTHTCPPCPA PELLGGPSVF LFPPKPKDTL MISRTPEVTC VVVDVSHEDP EVKFNWYVDG VEVHNAKTKP REEQYNSTYR VVSVLTVLHQ DWLNGKEYKC KVSNKALPAP IEKTISKAKG QPREPQVYTL PPSRDELTKN QVSLTCLVKG FYPSDIAVEW ESNGQPENNY KTTPPVLDSD GSFFLYSKLT VDKSRWQQGN VFSCSVMHEA LHNHYTQKSL SLSPGKHHHH HH
Product Science Overview
CD40 is a type I transmembrane protein that consists of an extracellular domain, a single transmembrane helix, and a cytoplasmic tail. The extracellular domain is responsible for binding to its ligand, CD40L (CD154), which is primarily expressed on activated T cells . The interaction between CD40 and CD40L is crucial for the activation of immune responses.
The recombinant human CD40 protein is typically expressed in HEK 293 cells (Human Embryonic Kidney 293 cells), which are widely used in biotechnology for the production of recombinant proteins. The recombinant form of CD40 is often produced as a fragment, usually encompassing the extracellular domain, to facilitate its use in various assays and research applications .
CD40 plays a pivotal role in the immune system by regulating the activation and differentiation of B cells, macrophages, and dendritic cells. Upon binding to CD40L, CD40 transduces signals that lead to the activation of several downstream pathways, including the NF-κB, MAPK, and PI3K pathways . These signaling cascades result in the production of cytokines, upregulation of co-stimulatory molecules, and enhancement of antigen presentation.
In B cells, CD40 signaling is essential for the formation of germinal centers, class-switch recombination, and the production of high-affinity antibodies. In macrophages and dendritic cells, CD40 activation enhances the production of pro-inflammatory cytokines and promotes the maturation and activation of these cells .
Recombinant human CD40 protein expressed in HEK 293 cells is widely used in research to study the CD40-CD40L interaction and its role in immune responses. It is also employed in the development of therapeutic strategies for various diseases, including cancer, autoimmune disorders, and infectious diseases .
For instance, targeting the CD40-CD40L pathway has shown promise in cancer immunotherapy, where activating CD40 can enhance the anti-tumor immune response. Additionally, modulating CD40 signaling is being explored as a potential treatment for autoimmune diseases by regulating the activity of autoreactive B cells and T cells .
