Anti-CD19 CAR mRNA-LNP
CD19 is a well-known biomarker expressed on most normal and malignant B cells and used as a target antigen found in B cell malignancies. CD19, also known as B lymphocyte antigen CD19, B lymphocyte surface antigen B4, T cell surface antigen Leu-12, and CVID3, is a transmembrane protein encoded by the gene CD19 in humans. This 95 kd transmembrane glycoprotein belongs to the immunoglobulin superfamily with two extracellular C2 group Ig-like domains and a relatively large cytoplasmic tail that is highly conserved across mammalian species. CD19 is expressed on all normal, mitogen-stimulated, and malignant B cells, with the exception of plasma cells. CD19 is ubiquitously expressed at all stages of B cell development and its surface expression is highly regulated. Because it is ubiquitously present on all B cells, it can serve as a B cell marker and a target for immunotherapy against neoplastic lymphocytes. This product is designed as a tool for the delivery and expression of anti-CD19-41BB-CD3 CAR mRNA for research. The product leverages the lipid nanoparticle (LNP) technology platform for simple and efficient delivery of anti-CD19-41BB-CD3 mRNA to a variety of mammalian cells in vitro and in vivo. The LNPs used are formulated with SM-102, DSPC, cholesterol and DMG-PEG2000 at an optimal molar concentration for a high rate of encapsulation and efficient mRNA delivery. The anti-CD19-41BB-CD3 CAR in this product is approximately 56 kD and consists of anti-CD19 scFv (single-chain variable fragment) linked to a second-generation CAR (chimeric antigen receptor) containing the CD8 hinge and transmembrane domain and 4-1BB and CD3ζ signaling domains. The full-length amino acid sequence of this anti-CD19-41BB-CD3 CAR mRNA-LNP product is available upon request.
mRNA-LNPs suspended in PBS (-Ca, -Mg) (pH: 7.0-7.4).
Cell Type Tested
T cells, HEK293S
Application & Handing
Upon receiving product, briefly pulse spin before opening to ensure product is at bottom of container. It is important not to spin for too long as this may rupture mRNA-LNPs. Do not vortex. Work with mRNA-LNPs on ice and minimize the time that the product spends at room temperature. After handling the product during experiments, return immediately to ice. mRNA-LNP products should only be handled with certified RNase-free reagents and consumables. Use of filtered pipette tips is highly recommended.
Anti-Mouse-CD19 CAR Expression in HEK293s Cells
Figure 1: CD19 CAR expression in HEK293S cells transfected by PM-LNP-0016, checked by FACS 48 hours post transfection. Cells were stained by 1° ab: Jackson ImmunoResearch Biotin-SP (long spacer) AffiniPure F(ab’)2 Fragment Goat Anti-Mouse IgG, F(ab’)2 fragment specific; 2° ab: Biolegend PE-Streptavidin. Transfected cells were gated by non-transfected HEK 293S cells grown under identical conditions. Cell morphology may be affected after transfection.
For Research Use Only. Not for use in diagnostic procedures.