Active Recombinant Human TNFRSF17(Met 1- Ala 54), Fc/His tagged

• Cat.No.: TNFRSF17-2348H
• Product Overview: Recombinant Human TNFRSF17(Met 1- Ala 54)(NP_001183.2) was expressed in HEK293, fused with a polyhistidine-tagged Fc region of human IgG1 at the C-terminus and a signal peptide at the N-terminus.

Specification

• Source: HEK293
• Species: Human
• Tag: C-Fc&N-His
• Protein length: Met 1- Ala 54
• Form: Liquid in sterile PBS, pH7.4.
• Bio-activity: Measured by its binding ability in a functional ELISA. Immobilized recombinant human BAFF at 1 μg/ml (100 μl/well) can bind human TNFRSF17. The EC50 of human TNFRSF17 is 0.0059 μg/ml.
• Molecular Mass: The recombinant human TNFRSF17/Fc is a disulfide-linked homodimer. The reduced monomer comprises 302 amino acids after removal of the signal peptide and has a predicted molecular mass of 34 kDa.
• Endotoxin: <1EU/ug by LAL method.
• Purity: Greater than 95% as determined by reducing SDS-PAGE.
• Storage: Store it under sterile conditions at -20°C ~-70°C. It is recommended that the protein be aliquoted for optimal storage. Avoid repeated freeze-thaw cycles.
• Reconstitution: It is recommended that sterile water be added to the vial to prepare a stock solution of 0.2 ug/ul. Centrifuge the vial at 4°C before opening to recover the entire contents.

Gene Information

• Gene Name: TNFRSF17 TNF receptor superfamily member 17 [ Homo sapiens (human) ]
• Official Symbol: TNFRSF17
• Synonyms: BCM; BCMA; CD269; TNFRSF13A; B cell maturation antigen; B-cell maturation factor; B-cell maturation protein
• Gene ID: 608
• mRNA Refseq: NM_001192
• Protein Refseq: NP_001183
• UniProt ID: Q02223

Citation

Preclinical evaluation of CD8+ anti-BCMA mRNA CAR T-cells for treatment of multiple myeloma

Journal: Leukemia    PubMed ID: 32632095    Data: 2020/6/26

Authors: Liang Lin, Shih-Feng Cho, Yu-Tzu Tai

Article Snippet:Viability of Descartes-08 cells was assessed using Acridine Orange and Propidium Iodide staining on the K2 Cellometer (Nexcelom Biosciences).Viability of Descartes-08 cells was assessed using Acridine Orange and Propidium Iodide staining on the K2 Cellometer (Nexcelom Biosciences).. CAR expression was determined by incubating CAR T-cells with 0.4 μg/ml of PE-conjugated recombinant BCMA (Recombinant human BCMA/TNFRSF17 protein, Fc/His-tagged, R-PE labeled; Creative BioMart, Shirley, NY) for 20 minutes at room temperature in FACS Buffer (DPBS, 0.5% BSA, 0.01% Sodium Azide).. Propidium Iodide was added at 1 μg/ml to stain non-viable cells.Propidium Iodide was added at 1 μg/ml to stain non-viable cells.

a Schematic representation of in vitro transcribed mRNA encoding anti-BCMA-CAR (Descartes-08). b The CD8+ T cells were activated and then transfected with mRNA encoding BCMA CAR for 4h, followed by flow cytometry (FCM) analysis to evaluate the percentages of BCMA-CAR on Descartes-08 and the paired control (ctrl) CD8+ T cells. Shown are histograms from a representative transfection among three experiments using three healthy donor T cells. c Frozen Descartes-08 cells generated via mRNA transfection (n=3) were thawed and the expression of BCMA CAR was then evaluated by FCM analysis from 4 hours (4h) to 12 day (12d). Data are shown as means ± SDs (error bars) from three independent experiments using three healthy donors, with each condition in triplicate. * P <.05, *** P <.001, **** P <.0001

NSG mice receiving MM1S-luc cells (2 × 10 6 cells per mouse i.v.) at day 0 were randomized into treatment groups (n=4 per group) on day 6. On days 7, 14, 21 and 28, T-cells were thawed from cryovials, washed, and resuspended for administration via i.v. injection with vehicle, 20 × 10 6 mock-transfected control TCR knockout (KO) CD8+ (ctrl CD8+) or Descartes-08 cells (showing 67% BCMA CAR expression by FCM analysis). On day 13, 20 and 27, all mice were treated with 60 mg/kg cyclophosphamide. a Bioluminescence (BLI) images (MM1S-fluc growth) of mice in each group on day 27 (after 3 doses of Descartes-08 cells or controls). Tumor growth ( b, means ± SDs) and survival ( c ) of animals are shown through timeframe of study. b - c Each of the 4 dosing days was marked with an arrow. Median survival days are 43 (vehicle), 44 (ctrl CD8+ T cells) and 69 (Descartes-08 CAR T) days. Survival curves of TCR KO Descartes-08 (Descartes-08) was significantly different ( P <.0001) compared with paired control TCR KO CD8+ T cells (ctrl CD8+ T) by log-rank (Mantel-Cox) test. Tumor growth curves were analyzed by 2-way ANOVA after log-transformation of data. Survival curve in animal model was plotted using the Kaplan-Meier method and compared by the log-rank test. **** P <.0001