GS Pressure Selection Technology

During the mammalian expression process, an effective way to enhance the cell line productivity is to amplify the copy of YFG (Your Favorite Gene) in the host cell. Creative BioMart offers MSX/GS pressure selection strategy to facilities the YFG amplification, which is specially combined with non-secreting murine myeloma cells (NS0) users. Additionally, this pressure selection strategy is also functional for the CHO cells.

GS-Pressure-Selection-Technology

Features of NS0 Cell

NS0 cell line has been commonly applied in the biotechnology industry. As a well-established mammalian protein expression host cell, NS0 is frequently used to generate hybridoma cells as a fusion partner to produce monoclonal antibodies. Currently, at least 6 commercial antibodies has been reportedly produced using NS0 cell line. NS0 cells applied in pharmaceutical production have proven to have many advantages:

GS-Pressure-Selection-Technology

Low endogenous glutamine synthetase level thus especially suitable for Glutamine Synthetase/Methionine Sulfoximine (GS/MSX) selection system.
Demonstrated high protein expression similar to CHO cell production.
Issues related to glycosylation.
Some NS0 lines require supplements (lipids) for high titers.

Technology Description

Glutamine synthetase is the only pathway to synthesize glutamine in mammalian cells. Some low endogenous GS level cell lines like NS0 and CHO require exogenous glutamine to survive in a glutamine-free medium. Transfecting of GS minigene to host cells allows glutamines synthesis while Methionine sulfoximine (MSX) can inhibit the synthesis process.

GS-Pressure-Selection-Technology

Based on the mechanism above, we use an expression vector encoding YFG (your favorite gene) + GS gene to transfect host cells, using an increased dose of MSX to kill the non-transfected or low-copy cells. As a result, only cell clones with a high YFG-GS copy will be able to survive.

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Features of Creative BioMart GS Pressure Selection Technology

Selection is based on glutamine synthetase, easy to be operated
No need for rounds of gene amplification
High yield without amplification and >5 g/L can be achieved
Suitable for both CHO and NS0 cell lines
Suitable for suspension growth in serum-free, chemically defined media
Expression procedure is fully optimized
High probability of monoclonality

Service Content and Timeline

Service Name	Service Description	Timeline
Gene and Vector preparation	Sequence design and codon optimization Gene synthesis Clone	2-4 weeks
Stable Cell Pool Selection	Transfection Stable cell pool screening Stable cell pool detection Selection of top 3 cell pools	6-8 weeks
Single Cell Cloning and Limiting Dilution	Amplify YFG with an increasing dose of MSX Screening single clone detection Subcloning for single subclone selection	4 weeks
Batch Culture or Fed-batch Culture	Selection top 10-30 clones based on expression profile	3-4 weeks
Cell Line Stability Evaluation	Select top 5-10 clones based on expression profile, for stability test, 10 generations	2 weeks
Research Cell Bank Preparation	Preparation of frozen cells from top clones for delivery	2 weeks
Scale-up Protein or rAb Production	Scale-up and purification of rAb or protein according to customers request	Variable based on request

GS-Pressure-Selection-Technology

Customers who need our assistance are more than welcome to contact us or submit an online inquiry. Our professional team will provide you with detailed guidance and solution ASAP. If you have any special needs or are interested in learning more about Creative BioMart's protein expression services, please feel free to inquire about more details.

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