High versatility. CelliMax® CHO P02 Medium is adapted to multiple cultivation processes, covering the entire workflow from cell thawing and passaging to transient transfection, fed-batch cultivation. No medium change is required for the entire duration of culture until harvest.

No, substitution of components within the kit is not recommended. To achieve maximum protein titer, users are recommended to use all components supplied in the kit. Substitution of any component may impact cell growth and reduce protein yield.

Option 1 has a wide range of applicable scenarios, featuring a straightforward workflow and enabling cultivation scale-up from shake flasks to bioreactors.

Option 2 introduces an additional centrifugation step on the day of transfection (D0), which can further improve the protein expression levels.

Two key requirements are recommended:

• Low-endotoxin or endotoxin-free plasmid DNA

Transient transfection is highly sensitive to endotoxins, which can induce cellular stress and apoptosis, reduce cell viability, and directly decrease protein yield.

• A plasmid DNA concentration of >800 ng/μL

Impurities such as RNA, proteins, and residual salts from plasmid preparation can interfere with the binding between DNA and the transfection reagent, impair complex formation, and reduce plasmid delivery efficiency into cells. In general, plasmid DNA concentration is positively correlated with protein yield.

To achieve optimal performance with CelliMax® TransCHO Kit02, high-purity, high-concentration plasmid DNA is highly recommended.

The recommended plasmid DNA concentration is 4.0 μg/mL.

For poorly expressing plasmids, the plasmid DNA concentration may be increased by 1.0–2.0 μg/mL, with the corresponding CHO Trans reagent02 increased to 8.5 μL/mL.

At high cell density during transient transfection, a temperature shift helps maintain high cell viability and extend culture duration, leading to improve protein expression levels.

Normally, no additional supplementation is required. CHO P02 Medium and uni FM03 Medium supplied with the kit contain stabilized glutamine, eliminating the need to prepare and add fresh glutamine.

When storage exceeds 6 months before usage, 2–4 mM L‑glutamine may be supplemented into CHO P02 Medium prior to use.

In general, the glucose addition level specified in the instruction is sufficient to meet cellular demands.

When the peak cell density exceeds 30 × 10⁶ cells/mL, an additional 2 g/L glucose is recommended based on the feed strategy.

If equipment is available to measure glucose concentration in the culture medium, monitor and supplement glucose concentrate daily or every other day to keep residual glucose ≥3 g/L.

The optimal harvest time varies depending on the CHO cell line and target protein.

Cells transfected with CelliMax® TransCHO Kit02 maintain high viability post-transfection, supporting a culture duration and protein yield comparable to those achieved in fed-batch cultures. In general, protein titer increases with extended culture time.

For optimal protein quality, harvest at Viability > 60% or Days 14~15 post-transfection.