Use of cell affinity chromatography for separation of lymphocyte subpopulations

Abstract

We describe the use of affinity chromatography for separation of cell populations that do not differ significantly with respect to gross physical properties such as size, density, or charge. Cell affinity chromatography exploits differences in cell surface macromolecules by passage of mixtures of cell populations through a column containing beads to which are attached chemical ligands with specific binding affinity for particular cell surface receptors. In this article we focus on the application of this concept to separation of mature T lymphocytes from peripheral blood. This serves as a model for the separation of these cells from bone marrow in order to prevent graft-vs.-host disease in bone marrow transplantation. However, the concept of cell affinity chromatography should find more general widespread utility in a variety of biotechnological applications. Thus, we introduce a simple theoretical framework which is necessary in order to understand the results that might be expected in any given situation. Finally, we use this theory to provide a tentative explanation for experimental observation of the effects of temperature and flowrate on the degree of separation achieved for our current pplication.