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Gibco™ ExpiCHO™ Stable Production Medium

Product Code. 16294262
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Product Code. 16294262 Supplier Gibco™ Supplier No. A3711001

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Description

Gibco ExpiCHO Stable Production Medium (SPM) is specifically designed to simplify your cell line development process into production with minimal effort and a high degree of confidence.

This chemically defined, protein-free, animal origin component-free formulation is fully optimized for large-scale fed-batch culture with ExpiCHO-S cells. ExpiCHO SPM supports an easy transition from transient to stable production, provides high titers without the need for additional medium optimization (see figures below), and works well with existing EfficientFeed C+ supplement.

ExpiCHO SPM is part of the ExpiCHO family, an end-to-end solution that empowers you to carry out your entire workflow from transient to stable expression without having to screen or optimize media and feeds.

Hassle-free scale-up
Simplify your process as you transition into stable clones with ExpiCHO SPM—no cell adaptation, no media optimization, no changes in cell doubling time and peak density. ExpiCHO SPM eases the transition using ExpiCHO-S cells and provides optimal performance with less need for fine-tuning from development scientists to get you to the next step faster. This medium supports high titers under standard fed-batch process conditions in conjunction with EfficientFeed C+ supplement. It is available in liquid format (this page) or in our AGT dry format from 10 L through 450 L equivalents with larger sizes available upon request.

ONE Solution
The ExpiCHO family includes products that are engineered to work seamlessly with each other. ExpiCHO SPM is a member of this family and allows expanded support of bioproduction from transient to stable protein expression. Look for ExpiCHO-branded protocols and methods to get optimal performance in the development process.

Members of the ExpiCHO family include:
• ExpiCHO Stable Production Medium (this page)
• ExpiCHO Expression Medium
• ExpiCHO-S cells
• Expifectamine CHO Transfection Kit

The power to streamline transfer to manufacturing
ExpiCHO SPM liquid format and AGT dry format demonstrate comparable cell growth and titer performance (see figure below). Therefore, scale-up and manufacturing using AGT dry format is simplified by knowing that the reconstituted AGT dry format will behave similarly to the liquid format used prior to scale-up. The AGT dry format is easy to reconstitute; just add water and it will automatically adjust pH and osmolality. This can help provide efficiencies in supply chain complexity, reconstitution complexity, and, therefore, overall expense.

TRUSTED_SUSTAINABILITY

Specifications

Cell Line ExpiCHO-S
Classification Animal Origin-free, Chemically-defined, Protein-free
Form Liquid
Product Type Stable Production Medium (SPM)
Product Line ExpiCHO
Quantity 1000 mL
Shipping Condition Room Temperature
Format Bottle
Content And Storage Store in refrigerator (2°C to 8°C).

Frequently Asked Questions (FAQs)

When subculturing ExpiCHO the day before a transfection to 3-4 x 10E6/mL, would you recommend pelleting the cells and resuspending them in completely fresh media, or simply replacing some of the media to split them down to the desired cell density?

Before transfecting ExpiCHO cells, there is no need to change out the media during the subculturing. This would become expensive and the conditioned media is not necessary.
For how many passages would you recommend using ExpiCHO cells before replacing them with new cells that were frozen at earlier passages?

For optimal performance with respect to protein production, we recommend thawing ExpiCHO cells and letting them grow for 2-3 passages prior to transfection. Overall, we do not recommend using the cells beyond ˜20 passages.

Is the ExpiCHO Stable Production Medium (SPM) compatible with the GS/MSX (Glutamine Synthetase Expression Technology) selection?

Yes, ExpiCHO Stable Production Medium (SPM) is compatible with the GS/MSX (Glutamine Synthetase Expression Technology) selection.
How much time do I need to consider for the stability study using ExpiCHO Stable Production Medium (SPM)?

After having obtained best expressing clones, a stability study is done. The time for a stability study varies (it depends how fast the cells double). It takes 60 generations or two months (whichever comes first).

When performing stable protein production using ExpiCHO Stable Production Medium (SPM), how is a stability study performed?

A stability study involves testing of ExpiCHO-S clones for 60 generations (60 population doublings) to see if high protein expression is maintained.

Is the transfection step different when stable selection is desired and ExpiCHO Stable Production Medium (SPM) is part of the workflow?

Yes, when stable selection is desired, transfection takes place in the presence of ExpiCHO Expression Medium but in the absence of the enhancer and feed. About 48 hours post-transfection, stable selection starts by addition of the appropriate selection antibiotics.
When should I not use ExpiCHO Stable Production Medium (SPM)?

We don't recommend using ExpiCHO SPM when you have an existing platform cell-line or feeds and are looking for a broadly usable medium, or when you need a transfection medium. We recommend using ExpiCHO Expression Medium for transfection.
When should I use ExpiCHO Stable Production Medium (SPM)?

We recommend using ExpiCHO SPM when transfection of ExpiCHO-S cells has been already completed with ExpiCHO Expression Medium, and you want to develop stable clones and scale up protein production.

What vector do you recommend using with ExpiCHO Stable Production Medium (SPM)?

We recommend using our pcDNA3.3-TOPO TA and pcDNA3.4-TOPO TA vectors. These vectors have only a single gene integration site so some modification will be necessary for antibody applications. You can use our GeneArt Express Cloning Service (https://www.thermofisher.com/us/en/home/life-science/cloning/gene-synthesis/geneart-plasmid-services/geneart-subcloning-service.html#express) to create a modification that will allow you to clone both the heavy and light chains into the vector (ideally joined by an IRES site). Please note that other vectors may also be used, and that you may want to test initially with 2-3 vectors for best results.

What vector did you use to produce the protein expression data with ExpiCHO Stable Production Medium (SPM)?

In our initial work, we used a proprietary vector that cannot be disclosed.

What is the difference between ExpiCHO Expression Medium and ExpiCHO Stable Production Medium (SPM)?

Both products are similar, however, formulation changes had to be made to make the ExpiCHO Stable Production Medium (SPM) scalable to AGT format. One such change is that it requires the addition of Glutamax whereas ExpiCHO Expression Medium does not. Additionally, ExpiCHO SPM is not compatible with transfection whereas ExpiCHO Expression Medium is compatible with transfection.
What is the intended use statement for ExpiCHO Stable Production Medium (SPM)?

The intended use statement for ExpiCHO Stable Production Medium (SPM) is as foll:
For Research Use or Further Manufacturing. Not for diagnostic use or direct administration into humans or animals.

Does ExpiCHO Stable Production Medium (SPM) contain Glutamax or Glutamine?

No. ExpiCHO Stable Production Medium (SPM) needs to be supplemented with Glutamax or Glutamine.
Can I perform transient transfection with ExpiCHO Stable Production Medium (SPM)?

No. Unlike ExpiCHO Expression Medium, ExpiCHO Stable Production Medium (SPM) is not recommended for transfection as it does not support high transient transfection efficiency.
What is ExpiCHO Stable Production Medium (SPM)?

ExpiCHO Stable Production Medium (SPM) is a chemically defined protein-free, animal origin component-free medium developed specifically to support high titer expression of stable ExpiCHO-S clones in suspension. The medium is designed to provide a seamless scale-up solution for customers using the ExpiCHO Expression System for transient production looking to transition to the development of stable ExpiCHO clones. The medium is formulated without hypoxanthine and thymidine for use in dihydrofolate reductase (DHFR)-amplified systems, without L-glutamine or GlutaMAX I Supplement for use in glutamine synthetase systems, and without phenol red in order to minimize the estrogen-like effects of phenol red.

ExpiCHO SPM allows for direct transition from transient expression to the large scale production of stable clones with no adaptation. You can start process development faster, and streamline or simplify transfer to manufacturing scale. ExpiCHO SPM is available in liquid format (Cat. No. A3711001) or in dry AGT format (Cat. Nos. A3711101, A3711102, and A3711103)

Note: ExpiCHO SPM is not compatible for use as a medium during the transfection step.
Can ExpiCHO-S Cells be used to create stable clones?

Our ExpiCHO-S Cells are derived from our cGMP banked CHO-S cells (Cat. No. A1155701), thus stable CHO-S selection methods are applicable to these cells. We recommend you follow the protocol outlined in the User Bulletin for the Creation and Scale up of a stable cell ine using ExpiCHO Products (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0017764_CreatScaleup_StableCellExpiCHO_UB.pdf)

For Research Use Only. Not for use in diagnostic procedures.

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