I am trying to obtain antibodies from a hybridoma cell line called TIB-117. I am not aquainted with this cell line and do not now how much to expect, at which density I should seed them or how long I should wait until harvest.
In the instructions from ATCC they are said to be cultured at 10 000-50 000 cells/ml.
At what density should I set them to start a culture for producution?
Can I "reuse" the cells after harvest or do I have to expand cells again?
I assume that I ought to let them grow approximately 10 days before I harvest them?
I would appreciate if someone could give me some advice
Sorry. I do not use this cell line. We only do transient expression in CHO for small samples.
What ATCC says in it's instructions is that you should seed the cells at 10^4cells/ml and let them grow up to 5.10^5 cells/ml which is supposed to take 2-3 days and then passage again.
I would probably suggest, that you begin according to instructions and "play" with them to get familiar. (But first make some frozen cell banks - just in case). You could try to passage them several times, start a batch culture and observe viability and maybe check for concentration of product you are looking for (ELISA/ western blot). Or maybe you can also find some more information in which you are interested in references of ATCC web page to these cells.
I don't know specifically this cell line. But from my own experience, once you begin, you learn very fast, what your cells can handle and what is too much for them. So good luck!
First of all, you get acquitted with TI B cellline, then, like any other hybridoma you need to obtain the OD and standardise the protocol. Refer Alisa- Hybridoma technology.
For my work, hybridomas were cultured in RPMI-1640 containing FBS. The color of media is pink before used. when hybridomas were cultured and secreted the mAb.... the media was changed from pink to yellow... if you found media is yellow... means you can collect the supernatant and change to new media... the cultured supernatant will be purified using Ammonium sulfate precipitation and follow by column. You can harvest hybridoma by pipetting and centrifuge. you can expand cell by transfering hybridomas into new flask with new media... or can freeze cell using 10%DMSO-RPMP. Pls aseptic for all ... dont make it contaminated... The cell concentration might not affect much, you can see under microscope that many or not.
Good luck
Surachai
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