SF Cell Line 4D-Nucleofector® LV Kit XL
What You May Also Need
Product Overview
The SF Cell Line 4D-NucleofectorTM LV Kit XL* is suited for larger scale transfection of cell lines in the LV NucleocuvetteTM Cartridge format when working with the 4D-NucleofectorTM LV Unit. With the LV NucleocuvetteTM Cartridge a variable volume of up to 20 mL can be transfected. The cartridge is automatically filled and emptied with help of the supplied input and output reservoirs. Alternatively bags can be used. An optional second input allows for separate feeding of mRNA to avoid its degradation.
This kit has been determined to be optimal for transfecting following cell lines and many others that use Solution SF as optimal NucleofectorTM Solution:
- HEK293 suspension clones
- CHO-S suspension clones
- K-562
- U-937
The respective cell type-specific Optimized Protocols or recommendations are available under "Instructions" or in our knowledge database. In these Optimized Protocols the best Nucleofection conditions are indicated. In addition, we share our experience and knowledge for treatment of individual cell types.
*Note: From the three cell line solutions (SE, SF, SG) that come with the 4D-NucleofectorTM System only SF is available for the large-scale formats so far. Solution SF is suited for the vast majority of cell lines that might be used in larger scale applications (e.g. suspension CHO or 293 cells). For other solutions please contact our Scientific Support.
Benefits
- Closed Nucleofection of up to 109 cells
- Conditions transferable from existing smaller scale protocols
- Automatic processing via tubing system
- Transfection of up to 20 mL reaction volume in less than 10 minutes
Applications
- For large scale transfection of up to 1 x 109 cells
- Transient production of potential therapeutic proteins or antibodies for construct screening
- Generation of large numbers of transiently modified cell lines for cell-based assays
Content & Storage
Content
22.5 mL SF Cell Line NucleofectorTM Solution
5 mL Supplement 1
1 x LV NucleocuvetteTM Cartridge
2 x 4D-NucleofectorTM LV Reservoir
Instructions
Safety Data Sheets (SDS)
Certificate of Analysis (CoA)
Please enter Lot Number, including all zeros, located on the product label and please take into account that it is case sensitive.
Lonza GS Xceed™ Cells transfected in small or larger scale show comparable IgG4 titers
For a human IgG4 antibody, similar titers of about 100 µg/mL were achieved in small scale (100 µl) and large scale transfections (1 ml fixed cartridge or 10 mL in flow through LV cartridge) in a 6-day batch culture without any feeds. IgG titers were increased by factor 2.5 when using higher seeding densities (not shown here). Switching to a 14-day fed-batch culture led to titers > 1 mg/mL (not shown here).
Transferability of Nucleofection® Conditions from Small to Large Scale
Expression of maxGFP® Protein in cells transfected in small volume (100 μL Nucleocuvette® Vessels) or larger volumes (1 mL or LV Nucleocuvette® Cartridge). Data represent the mean of various independent experiments.
