Hybridoma Development to create custom monoclonal antibodies is a multiphase process and with all the new technologies available to the scientist, the overall process might be a little overwhelming. Although hybridoma development cannot really be described as a black art, there are several important steps that can really impact the outcome of your monoclonal antibody production and even though the process of PEG fusion is fairly straight forward, imposing a common sense approach can make a difference.
Here are some suggestions:
Garbage in, garbage out. In order to acquire healthy, happy hybridomas, you need to start with healthy, happy splenocytes and myelomas. Harvest the splenocytes as gently as possible, and ensure myeloma partners are growing in log phase for fusion.
Plating strategies can be key to successful screening and clonal selection. Consider plating cell fusions from high-titer animals in an increased number of plates to help ensure hybridoma cells are at lower density and therefore unique clones will not be lost by overgrowth.
Once you have selected your hybridoma clones and are ready to scale up culture, treat your cells to some supplements in their culture media; the addition of cobalt, molybdenum, insulin, transferrin and selenium can all have a positive impact on cell viability and monoclonal antibody production.
Ever since hybridoma technology was discovered in the 1970’s, scientists have been tasked with improvement in the hybridoma development process and utilizing new technologies to do so. Hopefully using these suggestions will help you to create a successful batch of hybridomas to screen.