Clipos™ ANTI-CD19 mRNA Lipid Nanoparticles

Catalog:CDPM07

Unit Size:200 μL

INQUIRY Datasheet

Specifications
Description This product utilizes the lipid nanoparticle (LNP) technology platform for simple and efficient delivery of the Anti-CD19 mRNA to a variety of mammalian cells, including cell cultures which may typically be prone to difficulties in expressing desired proteins. The lipid nanoparticles used are formulated with, SM-102, DSPC, cholesterol, and DMG-PEG2000 at optimal molar concentration for a high rate of encapsulation and efficient mRNA delivery. CD19 is a well known biomarker expressed on most normal and malignant B cells. It serves a vital role in establishing instrinsic signaling thesholds. Studies have proved this marker ideal for target due its manageability and non expression on hematopoietic stem cells. This mRNA-LNP transfects the ScFv (single-chain variable fragment) of anti-CD19 linked to a 2nd generation CAR (Chimeric Antigen Receptor) containing CD8 hinge and transmembrane domains, and the 4-1BB and CD3ζ signaling domains. mRNA encapsulated in lipid nanoparticle, primarily in PBS (-ca, -mg). Dilute levels of: sodium acetate, ethanol, free mRNA & free lipids.
Lipid composition SM-102/DSPC/cholesterol/DMG-PEG2000
Encoding mRNA xCD19 CAR
In Vitro QC (Cell Types) T cells, HEK293S
Technology Applications CAR-T
Storage Store at 4°C
Technical Notes Work with mRNA-LNP on ice. It is important to minimize the time that the product spends at room temperature; after handling product during experiments, return immediately to ice.
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-LNP’s. Do not vortex.
mRNA-LNP products should only be handled with certified RNAase-free reagents and consumables; use of filtered pipette tips is highly recommended.
Usage 1. Prior to transfection, in a 12 well culture plate, plate your cells at [0.5E6 cells/mL] in a total of 1 mL per well. Ensure the cells you are using are viable and healthy. Try not to let your cells sit for longer than 5 minutes prior to transfection; cell clumping at the time of transfection may reduce transfection efficiency.
2. Briefly pipette mRNA-LNP solution up and down to resuspend. Add 20 uL of the mRNA-LNP mix dropwise directly to your 1 mL culture. Gently tilt plate back and forth to mix (not necessary if you are using cells which will be immediately placed back into a shaker). Place your transfected cells back into their original culture conditions.
3. Check cell expression by FACS at 24hr intervals after transfection.
*Note: This is a generalized protocol for transfection using mammalian cells. Transfection volume may be scaled down/up proportionately.
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