Isolation of B Cell Populations

This unit describes procedures for the isolation of pure human B cell populations. Two broad approaches to the isolation of pure human B cell populations are available, one based on techniques of negative selection and the other using positive selection. The negative selection approach has the advantage that the purified B cells obtained are less likely to be functionally altered by the isolation procedure. In addition, this approach does not require the use of expensive antibodies, which are usually the key element of a positive selection procedure. On the other hand, negative selection has the disadvantage that cell types not specifically targeted for removal (such as eosinophils) may be present in the final cell population. The positive selection method is an immunomagnetic positive selection using magnetic beads conjugated directly to an antibody specific for the B cell membrane determinant CD19. Also presented is a Support Protocol detailing the preparation of neuraminidase‐treated sheep red blood cells for use in rosetting.