ANTIBODY-PRODUCING CELLS: ANALYSIS AND PURIFICATION BY VELOCITY SEDIMENTATION

Velocity sedimentation cell separation is a simple and reproducible method for obtaining highly enriched populations of viable antibody-producing cells. Using suspensions of spleen cells prepared from mice immunized with sheep erythrocytes, fractions containing up to 2% 19S-PFC and 25% 7S-PFC can be obtained. Granulocytes constitute almost all of the remaining cells in these fractions. The sedimentation profile of 7S-PFC is very broad in comparison with that of cell populations known to be homogeneous in size (e.g. mouse erythrocytes). Analysis of the profile of 7S-PFC at different times after immunization suggests that the heterogeneity arises largely from the doubling in cell volume as a cell moves from one mitosis to the next. Early in the immune response, when the majority of the PFC are proliferating, the variation in sedimentation velocities is consistent with such a two-fold variation in cell volume. Late in the response, when most PFC have stopped proliferating, the sedimentation profile is more homogeneous. This analysis suggests that the fractionation procedure is sensitive enough to separate PFC according to their position in the cell cycle. Sedimentation velocities were also measured for several other classes of cells found in spleen. Comparison of these values shows that sedimentation velocity is a useful parameter for characterizing different types of cells.