Antibody production in vitro
Posted 27 November 2010 - 02:33 PM
Can anyone tell me how much antibodies I can expect from cultured hybridomas growing in bottle? I guess it is much less than producing Abs by ascitic liquid, right?
Posted 28 November 2010 - 12:16 AM
Posted 28 November 2010 - 04:06 AM
Depends very much on the cell line and the growing conditions but on average we get around 100 ug/ml in culture flasks.
Thanks a lot for your help regarding not only this last question but also previous ones!
You seem to know a lot about immunology! Please tell me: should I expect lower antibody levels if I culture the hybridomas in serum-free medium? Another question: you mentioned differences according to cell line, which are -in you view- the best lines to get highest Ab expression levels?
Posted 29 November 2010 - 04:17 AM
Posted 29 November 2010 - 05:29 AM
There are a number of smaller vessels for culturing hybridomas. I doubt that a PhD student or a post-doc will have access to a bioreactor. For scaling up we use a spinner cell, which can be incubated and spun in a normal incubator or a cell factory that produces roughly 10 mg/ml but these are expensive.
Biopharma with optimized cell lines in bioreactors are producing as much as 10mg/ml.
The net production in serum free medium is lower than with added serum. However, there are a number of advantages in growing in serum free medium, namely the absence of serum derived antibodies in your mAb prep. When I say it depends on the cell line, I mean that certain cell lines produce more than others intrinsically because of random chromosome loss during hybridoma stabilization. You can do very little about that. We, for example have had problems, when a highly producing cell was lost because it was overgrown by other less producing but faster dividing cells. That is why cloning after fusion is a must.