Separation of lymphocyte subpopulations by novel polymeric adsorbent with controlled micro-structure

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There is a stron demand for the materials which have the specific affinity to the particular cell population in the field or biomedical science and technology. The objective of this study is the molecular design of novel materials with specific affinity to lymphocyte subpopulations. We gave special attention to the diiferencial ionic character of cell surfaces owing to the differ- ences in the number and species of acidic functional groups expressed on their plasma membrane. Thus, basic amino groups were chosen as the main component of our designing polymeric materials to expect the specific interaction with B and T cells through ionic force. B and T cells are the two major subpopulations of lymphocytes, and their saparation is extremely important in the field of fundamental as well as clinical immunology.Through the systematic study on cellular interaction with a series of new multiphase material with polyuamine microdomains, polyHEMA-polyamine graft copolymer (HA) was found to adsorb B cells selectively out of a heterogeneous mixture of rat mesenteric lymphnode lymphocytes. As for the separation of B and T cells, the optical polyamine content in HA copolymer was 13wt% (HA13) for the polyamine grafts with Mn of 3750-6600 and is 7wt% (HA7) for that with Mn of 11000. Selectivity was lowered for HA copolymer having polyamine grafts with Mn of less than 1000 as sell as for ran- dom copolymer of HEMA with N,N'-diethyl-n-(4-vinylphenethyl)ethylenediamine.The most excellent result in terms of separation of B and T cells was obtained with HA graft copolymer with 13wt% of pollyamine grafts with Mn of 3600-6600. By using HA13(3750), T cell popu- lation with more than 92% purity was obtained in 95% yield as the column effluent. Also, by gentle pipetting, B cell population with 90% purity was recovered in 85% yield from the column. Consequently, our results demonstrated a promising feature of cell adsorption chromatography as a major tool for cell separation.