Notes on Making Rat x Y3 Monoclonal Antibody Producing Hybridomas

This note is not intended as a full protocol for making monoclonal antibodies, but is to give a few hints and tips for the use of Y3/Ag1.2.3. Generally speaking the methodology is similar to making murine (eg. with NS0) monoclonal antibodies, but there are a few critical differences which are commented on below:

1) Why we like to make monoclonal antibodies with Y3/Ag1.2.3

Although Y3/Ag1.2.3 has its own myeloma light chain, their are numerous characteristics that make this a better parent line for making hybridomas than the 'Y0' chain loss variants or using NS0 to make rat x mouse hybrids. These are:

2) Notes on immunization.

3) The fusion.

4) Chain loss and isotype switch variants

5) For full details see:

Clark, M.R., & Waldmann, H. Production of murine monoclonal antibodies in Methods in Hematology 13:1-20 (1986).

Steve Cobbold - last updated 27th July 1998.