Tag: <span>hybridoma culture</span>

In an attempt to express and isolate BoLA-NC3*00101, a bovine, non-classical, major histocompatibility complex (MHC) class I protein, the mouse mastocytoma cell line P815 was transfected with the NC3*00101 transgene. The transfected cells were checked for expression by flow cytometry and positive stable transfectants were sorted using a fluorescence-activated cell sorter (FACS). A large amount of purified protein is required for the generation of monoclonal antibodies in order to immunize the mice and then boost the host for several months to get highly-specific antibodies. To avoid using a large number of cell culture flasks and shorten the time required to obtain a sufficient quantity of protein, stably-transfected BoLA-NC3*00101-expressing cells were grown in a pitched-blade bioreactor. One week of culturing in the bioreactor yielded a large cell mass which was used to isolate and purify the protein…