Does anyone have experience with high level protein expression in mammalian cells like CHO? What is the best vector for this purpose?
Lonza Biologics has vectors for high level of expression in CHO cells and NSO cells.You can check in their website.
There are lots of options , some which are generally free to use others which have been developed by companies for commercial purposes for which you usually require a license. In general those which have been developed by companies and for which a license is required give you the best chance of obtaining high levels of expression e.g. IgGs at sevaral grams per liter. However the high levels of expression come from a combination of factors, vector, host, selection process and culture process. Is this something you wish to use to quickly generate material for research purposes using transient cell culture or something to develop a stable cell line for a production process?
Dear philip cunnah,
thank you for your comments. I know optimization in different level is needed for high level expression. Using of good vector is the first step! I meant development of stable cell line for a production process.
about LONZA vectors, is there commercially available??
Lonza vectors are commercially available. I think they are expensive.although i would suggest to check with them.Usually companies tend to buy them for production of biopharmaceuticals.Invitrogen also probably has some vectors for mammalian expression.
You might want to give the Freedom CHO S kit, Life Technologies (Invitrogen) a look. There is a relatively low license fee for research use which is included in the cost of the kit and you also have the added benefit of buying this directly , "off-the-shelf" from Life Technologies without having to go through a process of CDAs and license agreements which is generally the case for some of the other technologies around.
The eventual commercial license of this system, e.g. if you take something into clinical devlopment and beyond, will have to be negotiated with Life but is relatively low priced when compared to some of the other systems available.
In 4 - 5 months, after amplification and limiting dilution selection in combination with Life's platform fed batch culture process, you should be able to get to stable clone candidates with levels of expression in the 1 - 2 gram / litre range for IgGs .
With other technolgies you might be able to get higher levels in shorter time, depending on your resourses. If you have the time and cash to spend I would encourage you to consider comparing a few different technologies.
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