Extraction and Purification of Soluble HL-A Antigens from Exhausted Media of Human Lymphoid Cell Lines
Open Access
- 1 September 1973
- journal article
- Published by Oxford University Press (OUP) in The Journal of Immunology
- Vol. 111 (3) , 783-788
- https://doi.org/10.4049/jimmunol.111.3.783
Abstract
HL-A alloantigens could be isolated in soluble form by 3 M KCl extraction of membranous debris present in the exhausted, cellfree media of several cultured human lymphoid cell lines. These antigens were highly specific, possessed good immunologic potency, and could be obtained in high yield from this source material. Purification of HL-A antigens could be readily achieved by preparative acrylamide gel electrophoresis resulting in the isolation of an electrophorectically homogeneous moiety with a 20-fold increase in serologic reactivity. Yield and immunologic potency of HL-A antigens from exhausted culture media did not vary during different phases of the cell growth cycle in contrast to whole lymphoid cells where optimal yields were achieved in the late log or resting phase. The condition of the cell membrane, cell metabolism, and membrane turnover seemed to have an appreciable influence on the yield and immunologic potency of soluble antigens that can be extracted with 3 M KCl from cultured human lymphoid cells.Keywords
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