Immunomagnetic purification of T cell subpopulations.

This unit describes a general method for physical separation of cell subpopulations from a heterogeneous mixture of cell types. The technique relies on having an appropriate monoclonal antibody or mix of monoclonal antibodies that distinguish between the cell types being separated. Its advantages over other antibody-mediated selection techniques are purity of resulting cell preparation, reproducibility of separation, and ease of handling small to large numbers of cells (10(6) to 10(10) cells). The basic protocol outlines preparation of purified human T lymphocytes. The method produces good yields of pure T cells prepared by negative selection (i.e., not labeled with antibody). The alternate protocol applies similar principles for purifying CD4(+) T cells, and presents refinements of the basic protocol that minimize cost and improve convenience.