Mirus Bio receives ISO 13485:2016 certification, underscoring the quality of processes used to support GMP product portfolio. Read more

mirus bio logo

L-Glutamine Solution – 100 mL

High density growth and quick adaptation of suspension CHO cells

SKU MIR 6240 Categories ,

$43.00

Description

Ideal for Biotherapeutic Protein Production in Suspension CHO cells

CHOgro® Expression Medium is a chemically defined, hydrolysate-free and animal-origin-free medium. CHOgro® is formulated to provide high density cell growth, and many suspension CHO cells (e.g. Freestyle CHO-S) can easily and quickly grow in CHOgro® Expression Medium with minimal adaptation. CHOgro® Expression Medium requires supplementation with L-Glutamine and Poloxamer 188 before use.

CHOgro® Complex Formation Solution is an animal-origin-free solution specifically formulated for optimal transfection complex formation in conjunction with CHOgro® Expression Media and TransIT-PRO® Transfection Reagent to provide a completely animal-origin-free expression system.

  • High Density Growth – CHO-S cells grow > 12 x 106 cells/ml
  • Quick Adaptation – CHO-S cells double within 24hr of complete media change from FreeStyle™ Expression Media
  • No Cell Clumping Post-Transfection – Obtain accurate cell counts and high viability
  • Quality – CHOgro® Medium and Complex Formation Solution are manufactured under cGMP conditions in an ISO-compliant facility
  • Regulatory Friendly – ALL components are animal origin free

 

CHOgro® Expression Medium is also available in 10 L quantities in powder or liquid format. Please contact support to inquire: techsuppport@mirusbio.com

 

SKU: MIR 6240

Supporting Data

 

CHOgro® Expression Medium Yields Multi-fold Increases in Antibody Titer. Human IgG1 was produced by transient transfection using TransIT-PRO® (1:1), FreeStyle™ MAX (1.25:1.25)  or 25kDa linear PEI (6:1) transfection reagents according to the manufacturers’ or published protocol (reagent:DNA ratio). Transfections were performed using 1 µg plasmid DNA per milliliter of culture and cell densities of 2 x 106 cells/ml or 1x 106 cells/ml  for the CHOgro® Expression Medium (red bars) or FreeStyle™  Expression Medium (blue bars), respectively, at the time of transfection.  FreeStyle™ CHO-S cells were cultured in CHOgro® Expression Medium or FreeStyle™ CHO Expression Medium  and plated into non-treated 6-well plates (2ml/well) for transfection. Antibody levels were also analyzed from day 6 clarified supernatants using a human IgG ELISA (ZeptoMetrix). Error bars represent the standard deviation of triplicate technical replicates.

 

Suspension CHO Cells Grow to High Density in the CHOgro® Expression Medium. Triplicate flasks of FreeStyle™ CHO-S cells were seeded in CHOgro® Expression Medium (red line) or FreeStyle™ CHO Expression Medium (blue line) at cell density of 0.5 x 106 cells/ml, 40 ml per 125 ml shake flask (Thomson). Cell counts (solid line) and viability (propidium iodide staining, dotted line) were measured daily using a Guava® easyCyte™ 5HT flow cytometer (EMD Millipore).  Error bars represent the standard deviation of three readings of biological triplicates.

 

CHOgro® Media Exchange Leads to Higher Protein Production. FreeStyle™ CHO-S cells were cultured in FreeStyle™ CHO Expression Medium or CHOgro® Expression Medium.  Twenty four hours prior to transfection a subset of the cells grown in FreeStyle™ CHO Expression Medium were spun down and exchanged with 100% fresh CHOgro® Expression Medium.  The cells were allowed to grow and adapt for 24 hours prior to transfection with FreeStyle™ MAX (1.25:1.25) or TransIT-PRO® (1:1)  transfection reagents according to the manufacturers’ protocol (reagent:DNA ratio) and a human IgG1 encoding construct. Transfections were performed using 1 µg plasmid DNA per milliliter of culture and cell densities of 1 x 106 cells/ml for cells transfected with FreeStyle™ Max and 2 x 106 cells/ml for cells transfected with TransIT-PRO®. All cells were plated into non-treated 6-well plates (2ml/well) for transfection. (A) Workflow schematic of media exchange of CHO-S cells from FreeStyle™ CHO Expression Medium to CHOgro® Expression Medium (black arrow) or the normal CHOgro® Expression System (red arrow) (B) Day 6 supernatants were clarified and analyzed using a human IgG ELISA (ZeptoMetrix). Data is normalized to the complete CHOgro® Expression System (red bar).  Error bars represent the standard deviation of triplicate technical replicates.

 

Less Cell Clumping is Observed with the CHOgro® Expression System. FreeStyle™ CHO-S cells were cultured in  CHOgro® Expression Medium or FreeStyle™ CHO Expression Medium and seeded into a 125 ml shake flask (20ml culture volume, Thomson) for transfection. Human IgG1 was produced by transient transfection using TransIT-PRO® (1:1) or FreeStyle™ MAX (1.25:1.25) transfection reagents according to the manufacturers’ protocol (reagent:DNA ratio). Transfections were performed using 1 µg or 1.25 µg plasmid DNA per milliliter of culture and cell densities of 2 x 106 cells/ml  or 1x 106 cells/ml  for the CHOgro® or FreeStyle™  System, respectively, at the time of transfection. Pictures were taken of representative flasks and cells (inset) 6 days post-transfection.

Resources

Specifications

Storage Conditions
CHOgro® Expression Medium: Store at 4°C
CHOgro® Complex Formation Solution: Store at 4°C
Poloxamer 188 Solution, 10%: Store at room temperature
L-Glutamine Solution, 200 mM: Store at -20°C

Product Guarantee
All Configuration: As labeled on the product

Usage Statement
All Configurations: For Research Use Only.

Animal Origin Statement
All Configurations: This product is animal origin free.

 

Technical Product Literature

Quick Reference Protocols
CHOgro® Complex Formation Solution Quick Ref Protocol (PDF)
CHOgro® Medium Dry Powder Quick Ref Protocol (PDF)
CHOgro® Medium Quick Ref Protocol (PDF)

Product Data Sheets
L-Glutamine Solution Product Data Sheet (PDF)
Poloxamer 188 Product Data Sheet (PDF)

SDS
CHOgro® Complex Formation Solution SDS (PDF)
CHOgro® Medium Dry Powder SDS (PDF)
CHOgro® Medium SDS (PDF)
L-Glutamine Solution SDS (PDF)
Poloxamer 188 Solution SDS (PDF)

Additional Protocols
TransIT-PRO® for Expi293F™ Cells (PDF)
CHOgro® Expression System Full Protocol (PDF)
TransIT-PRO® Full Transfection Protocol (PDF)

 

Additional Resources

Poster: Development and Optimization of CHOgro® Transient Expression Technologies for High Titer Antibody Production in Suspension CHO cells (PDF)

FAQs

See the CHOgro FAQs

Using transfection reagents and enhancers from Mirus has given our platform a competitive advantage. Increasing efficiency and lowering costs for all of our porgrams.

 

Sally Mader, PhD
ZYZ Theraputics