Reinitiating a hybridoma culture from a frozen stock occasionally results in only a small number of viable hybridoma cells. Typically, this is caused by poor freezing technique, low viability of the hybridoma cells at the time of freezing, or improper maintenance of frozen cell stocks. It can be extremely difficult to propagate the few viable cells that survive. Using proprietary technology/medium developed by Cell Essentials, we have been successful in rescuing hybridomas where only a few (1-5) cells were viable. Cells are expanded and vials frozen back.
Antibody production from established hybridoma cell lines can diminish over time due to a decrease in either the number of cells that secrete antibody or the variations in the amount of antibody secreted. In many cases, antibody production can be improved by using two different techniques. These include the identification and isolation of an FACS enriched population of hybridomas producing the uppermost amount of antibody in the culture and/or classic single cell recloning of the hybridoma cell line.
- Enrichment – Multiple rounds of FACS enrichment are carried out and the top antibody producing cells are expanded and frozen.
- Single cell cloning – Antibody-positive clones as determined by antibody capture ELISA are isolated. The clone that exhibits good growth and produces the highest antibody titer is expanded and frozen.