Hybridoma - (Sep/18/2014 )

Hi all,

I am getting high sera titers in mice by protien immunization but not able to convert titers into secretory hybrids.Any suggestions?

Hi,

sometimes it can happen. In my experience, 1 every 30-35 fusions.

It may depend on the age of the mice, what I've observed is that if the immunized mouse is old, it gives relatively poor secreting hybridomas.

Alternatively it can depend from the myeloma cells, if thawed from a long time.  Moreover, no secretory hybrids can be a result of mycoplasma contamination. The last idea is that you can have a problem in the screening phase (like a secondary antibody or coating if you are using ELISA.

May I've just told you obvious explanations, I hope this can help you.

Ask me if you have questions!

Hi,

According to the indications of Deborah.

Another idea is to test the HGPRT enzyme deficiency in your mieloma cell line before cell fusión with splenocytes from hyperimmune mice.

One possible explanation may be that the mieloma cell line express again  (partially or completely) HGPRT enzyme, which many unfused myeloma or mieloma-mieloma hybridomas cells may grow, resulting in wells with cell growth in your plates, but many of these myeloma cells are not secreting antibody.

I hope this can help you