CAR-T Cell Therapy Target Protein

Two landmark events occurred for CAR-T cell therapy in 2017: two anti-CD19 CAR-T cells were approved by FDA for the therapy of acute lymphoblastic leukemia (ALL) and relapsed or refractory large B-cell lymphoma, respectively. From then, increasing number of researchers have been dedicated to CAR-T cell therapy research.
What is CAR-T cell therapy?It is a kind of immunotherapy mainly about isolating the patient's T cells and genetically engineering the cells to express chimeric antigen receptors (CARs), which can recognize a specific tumor associated antigen, such as CD19, BCMA and CD20. After infusing the engineered CAR-T cells into the body, the tumors cells are targeted efficiently.
Before infusing CAR-T cells into patients, CAR+% assay through flow cytometry is inevitable. The most commom method for CAR+% detection is using target protein as detection reagents. Sino Biological has successfully developed over 6000 recombinant proteins, covering most of the investigational CAR-T cell therapy targets. We have developed biotinylated proteins and fluorescent labeled proteins to simplify your research procedures. The proteins with high purity and high binding activity, have also been validated in high-class citations. Therefore, CAR-T cell therapy target proteins in Sino Biological Inc. are the first choice for your scientific research.

CAR-T Cell Therapy Target Proteins Available

Solid tumor targets

Official name Alias Diseases
CA9 CAIX metastatic renal cell carcinoma
CD274 PD-L1 glioblastoma
CD70   b-cell malignancies, pancreatic cancer, renal cell cancer, breast cancer, melanoma, ovarian cancer
CEACAM5 CEA a variety of solid tumors(lung cancer,colorectal cancer,gastric cancer, breast cancer and pancreatic cancer )
EGFR   colorectal cancer
EGFRvIII   glioblastoma
EPCAM CD326 hepatoma, gastric cancer
EPHA2   malignant glioma
ERBB2 HER2 a variety of solid tumors (breast cancer,ovarian cancer,lung cancer,gastric cancer, glioma, pancreatic cancer)
FAP   malignant pleural mesothelioma
FOLH1 PSMA prostate cancer
FOLR1 FRα ovarian cancer
GPC3 Glypican-3 hepatocellular carcinoma, squamous cell lung cancer
IL13RA2   glioma
KDR VEGFR2 metastatic carcinoma, metastatic melanoma, renal cancer
L1CAM CD171 ovarian cancer, neuroblastoma
MET c-MET breast cancer
MSLN Mesothelin mesothelioma, pancreatic cancer, ovarian cancer
Muc1 CA15-3 hepatocellular carcinoma, non-small cell lung cancer, pancreatic cancer, breast cancer
PROM1 CD133 liver cancer, pancreatic cancer, brain tumor, breast cancer, ovarian tumor, colorectal cancer, acute myeloid and lymphoid leukemias
ROR1   non-small cell lung cancer
ULBP1 NKG2D ligand 1 ovarian tumor, a variety of hematological malignancies
ULBP2 NKG2D ligand 2 ovarian tumor, a variety of hematological malignancies

Hematologic cancer Targets

Official name Alias Diseases
CD19   leukemia, lymphoma, multiple myeloma
CD22 Siglec-2 b-cell lymphoma
CD33 Siglec-3 myeloid malignancies
CD38   acute myeloma leukemia, multiple myeloma
CD5 Leu-1 t cell acute lymphoblastic lymphoma
CD7   acute myeloid and lymphoid leukemia
IL1RAP IL1R3 chronic myeloid leukemia
IL3RA CD123 myeloid malignancies
MS4A1 CD20 leukemia, lymphoma, multiple myeloma
NCAM1 CD56 acute myeloma leukemia
PROM1 CD133 liver cancer, pancreatic cancer, brain tumor, breast cancer, ovarian tumor, colorectal cancer, acute myeloid and lymphoid leukemias
ROR1   non-small cell lung cancer, breast cancer, leukemia
SDC1 CD138 multiple myeloma
TNFRSF17 BCMA multiple myeloma
TNFRSF8 CD30 lymphoma
ULBP1 NKG2D ligand 1 a variety of hematological malignancies, ovarian tumor
ULBP2 NKG2D ligand 2 a variety of hematological malignancies, ovarian tumor

Validated Application in Citations for CAR-T Cell Therapy Targets

CAR+ expression detection using recombinant CEA protein

Title: Phase I escalating-dose trial of CAR-T therapy targeting CEA+ metastatic colorectal cancers
Authors: Zhang C, et al.
Journal: Molecular Therapy

CAR expression were detected in CAR-T cells by FCM method using CEA protein (Cat. No. 11077-H08H). The median CAR+% is 33.7% and the range is 14.7%–43.2%.

Fig. 1 CAR expression were detected in CAR-T cells by FCM method using CEA protein (Cat. No. 11077-H08H). The median CAR+% is 33.7% and the range is 14.7%–43.2%.
PBMCs were activated by immobilized CD3 and CD28 antibodies. On day 2, T cells were infected with lentiviral vector in six-well plates with polybrene. After viral transduction, T cells were expanded by IL-2 for approximately 12–14 days to obtain enough cell dose. Quality control of cell products was performed by FCM. For the cell products, the CAR+ %, proportion of T cells (CD3+), NK cells (CD16+CD56+), NKT cells (CD3+CD16+CD56+), CD4/8 ratio, T memory cells, and checkpoint inhibitors (TIM-3, PD-1, LAG-3) on the T cell surface were measured on the day before CAR-T infusion. To ensure transfection, CAR+ % detection 4–5 days after viral transfection were also applied.

CAR+ expression detection using recombinant EGFR protein

Title: Chimeric antigen receptor
Authors: Shiku H, et al.
Journal: US 9,175,308 B2

CAR-positive rate was detected using Recombinant Human EGFR (from Sino Biological Inc.). The CAR-positive rate is relative to copy numbers of the retrovirus.

Fig. 2 CAR-positive rate was detected using Recombinant Human EGFR (from Sino Biological Inc.). The CAR-positive rate is relative to copy numbers of the retrovirus.
A human PBMC which had been collected after obtaining informed consent was infected two times with an undiluted solution or a 2-fold, 4-fold or 8-fold dilution of each virus solution to prepare a PBMC expressing anti-EGFR-CAR. Five days after the second virus infection, Recombinant Human EGFR in which the C-terminal end was His-tag-modified was added to the infected cells, and then a biotin-labeled anti-His-tag antibody was added. There after, the cells were stained with streptavidin-PE and a FITC labeled anti-Human CD8 antibody. Using a flow cytometer, the stained cells were subjected to measurement of a rate of PE-positive cells in FITC-positive cells, that is, a rate of cells positive for CAR that binds to EGFR in CD8-positive cells. Further, the infected cells were recovered 5 days after the second virus infection, and subjected to measurement of the number of copies of the virus incorporated into the genome.

CAR-T Cell Therapy Target Protein Advantages

High purity, most >95% as determined by SDS-PAGE

The purity of human BCMA protein (ECD, Fc Tag) (Cat. No. 10620-H38H) is >95% as determined by SDS-PAGE.

Fig. 3 The purity of human BCMA protein (ECD, Fc Tag) (Cat. No. 10620-H38H) is >95% as determined by SDS-PAGE.

The purity of human Mesothelin protein (Fc Tag) (Cat. No. 13128-H02H) is >99% as determined by SDS-PAGE.

Fig. 4 The purity of human Mesothelin protein (Fc Tag) (Cat. No. 13128-H02H) is >99% as determined by SDS-PAGE.

Validated high binding activity

Flow cytometric analysis of FITC-CD19 protein (Cat. No. 11880-H08H-F) on CD19-CAR transfected 293 cells

Fig. 5 Flow cytometric analysis of FITC-CD19 protein (Cat. No. 11880-H08H-F) on CD19-CAR transfected 293 cells.
 

The biotinylayed human Glypican-3 protein (Cat. No. 10777-H08H-B) and unconjugated human Glypican-3 protein (Cat. No. 10777-H08H) binds CD20 antibody drugs in a functional ELISA.

Fig. 6 The biotinylayed human Glypican-3 protein (Cat. No. 10777-H08H-B) and unconjugated human Glypican-3 protein (Cat. No. 10777-H08H) binds CD20 antibody drugs in a functional ELISA.

Low endotoxin: < 1.0 EU/μg by the LAL method

Biotinylated proteins and fluorescent labeled proteins available

• Sino Biological provide ready-to-use biotinylated proteins for most of the CAR-T cell therapy targets, all of which go through strict QC standard and show high activity. To learn more about biotinylated proteins.
• Fluorescent labeled proteins are also under quick research and will be in the sale soon!

Custom recombinant protein expression and purification service

If you can not find the protein you need, we also offer custom service for protein production. With five protein expression systems and over 10 years experience, we are professional in recombinant protein produciton. To find more……

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