An early pioneer in the area of disposable bioreactors, the WAVE Bioreactor was the first single-use bioreactor system when it was presented to the bioprocessing community in 1996. Since then, the range of products has expanded to include larger scale systems and advanced optical sensor and control technologies. WAVE Bioreactor systems are today widely used in both research and manufacturing operations. The latest WAVE bioreactor, ReadyToProcess WAVE 25, combines the ease-of use that comes with the rocking technology as such, with intelligent control and advanced sensor technology.
I am interested in a flexible perfusion setup to achieve high cell concentrations in insect cell culture. I am using SF 9 cells and would like to run initial experiments at small volumes of 1 to 3 L. Furthermore I am interested in an option for scale-up at a later time.
Perfusion cultures can be run using Cellbag bioreactors with integrated filter for cell retention. Harvest can be withdrawn directly through the filter, using a harvest line that ends in a luer connector at the bag wall. The filter has a nominal pore size of 7 µm and floats on top of the culture liquid. The … Continued
Are there flexible options for DO regulation in the WAVE bioreactor as I plan to run a mammalian cell culture under hypoxic conditions?
To maintain hypoxic culture conditions in the WAVE Bioreactor, it is possible to connect nitrogen instead of compressed air to the gas mixer. Total: 0 0 0 0
The WAVE Bioreactor can be used for the large scale cultivation of adherent cells and manufacture of viruses. We recently scaled a microcarrier based Vero cell culture from 10 L working volume in WAVE 20/50 to 50 L working volume in WAVE 200. Comparable cell growth was obtained at both working volumes and the virus … Continued
Can you tell me the main features the new WAVE bioreactor offers that make it better than the old WAVE system.
The new system was developed with a focus on reliability and ease of use. The controllers for culture parameters like temperature, pH and DO as well as the liquid handling have been updated. They allow for an intelligent process control, taking into account the Cellbag bioreactor size and the culture volume. If desired the user … Continued
In my lab we are working on producing antibody using hybridoma for both in-vivo studies and also pre-clinical work. We are using shake flask, but now need to produce more volume. I am wondering what system you would suggest for smaller scale production and if you would recommend perfusion culture or not.
Multiple hybridoma cell lines have been successfully cultivated in WAVE Bioreactor systems. Using different Cellbag bioreactors, cultivation volumes from < 0.5 L to 25 L can be run on the same instrument. After an initial characterization of the cultivation process in batch mode, a process optimization targeting perfusion is a viable option to increase the … Continued
Can you explain in a bit more detail how the internal floating filter works? It seems like an efficient set-up.
The floating filter is used to retain cells in the Cellbag Bioreactor during perfusion cultivation or medium exchange. It consists of filter material with a nominal pore size of 7 µm, which is laminated to a plastic film covering the back of the filter. Due to the lower density, the filter floats on top of … Continued
Wave bioreactor is a very useful choice for quicker monoclonal antibody production in small institutes. I have a feeling that some clones like “wave”, but some clones did not, espceially when the clone secrets IgG3. The cells either showed low viability, low growth rate or low antibody concentration. We also had bad luck when using wave bioreactor for CHO K1 cells. Can you tell us what physiodynamic mechanisms may influence cell growth of certain cells? Is there any way to control this?
There are certainly clonal differences in e.g. shear sensitivity or general robustness. The type of recombinant protein should not have a major impact except when expressing receptors or other surface proteins. As with other bioreactors, cells may need adaptation to the agitated environment in a WAVE Bioreactor. Taking special care about equilibrating the bioreactor before … Continued
I use shakers for transient expression with HEK, however the expression level is low and I have to increase the productivity. Will I get more protein in a WAVE? Do you have any other tip for me?
The efficiency of transient transfection is very much dependent on the method used for DNA transfer. Most commonly polyethylenimine (PEI) is used as transfection reagent. The optimum DNA:PEI ratio is dependent on the cultivation medium used and has to be established first. Also the maximum PEI concentration the cells tolerate and the requirement for media … Continued
We are looking into scale up of BHK cell cultures from shake flasks to a Wave and experience sluggish growth of the cells in the bioreactor. Do you have recommendations for optimizing the culture conditions?
When transferring a clone to a new cultivation system, it is important to make the most out of the characterization data generated in the previous one. E.g. what is the optimum seeding concentration, are the cells very sensitive to the cultivation pH, what is the limiting nutrient in the cultivation medium used, how long can … Continued
After transfer of cell culture medium into the Wave bag, the pH is more alkaline than the set-point. Switching on the pH control results in maximum CO2 output. On several instances the pH then dropped up to 0.3 units below the set-point and it took several hours until the pH reached the acceptable range and the cells could be transferred to the bag.
CO2 control of the pH in Cellbag Bioreactors occurs via the headspace. Depending on gas flow, bag size and agitation conditions some delay will occur until the gas composition in the headspace is exchanged and the CO2 has dissolved in the cultivation medium. Suggested gas flows and agitation conditions for specific Cellbag Bioreactor sizes can … Continued
We want to cultivate cholesterol dependent NS0 cells in a Wave reactor. It is said that cholesterol is adsorbed to the bag film. What can be done to achieve a sufficient concentration of the lipid in the culture?
The problem of cholesterol delivery to NS0 cells was investigated when differences in cell growth were observed in serum containing culture and serum free medium supplemented with lipids and synthetic cholesterol. The conclusion of the authors was that the interaction of the lipid carrier, in this case methyl-beta-cyclodextrin, with the bag film led to cholesterol … Continued
I saw a blog on the Cell Culture Dish that talked about using the Wave for cell banking for CHO cells. I was wondering if the same principal could be applied to cell banking of vero cells.
Answer: In principle a similar approach can also be applied to Vero cells. The difference is that the cells are adherent and require microcarriers for cultivation in the WAVE Bioreactor. A microcarrier often used for Vero cultivation is Cytodex, please see literature reference in my answer regarding the use of the WAVE Bioreactor for the … Continued