Further characterization and standardization of mouse monoclonal antibodies reacting with M/N blood group antigens.

The M/N-blood-group system is very suitable for investigating the reactivity of monoclonal antibodies because the antigenic structure seems to be very dependent on influences of the microenvironment, whereas on the other hand the biochemistry of these antigens is very well known. The distribution of these antigens within the population makes this system very interesting for forensic investigations and therefore one needs specific reagents with constant quality. We have immunized BALB/c mice with whole red blood cells and obtained a series of monoclonal antibodies. Some of these reacted with epitopes which are very dependent on pH, temperature and buffers concerning the binding of the antibodies and the specificity. By standardizing the monoclonal preparation in this respect it was possible to obtain specific reagents. In addition we found antibodies reacting with epitopes which were not influenced by the microenvironment. These antibodies always gave specific results. To get further criteria for standardization of these reagents we have adapted the method of flow cytometry for binding studies of monoclonal antibodies against red blood cell antigens.