Hydrophobic chromatography of cells: adsorption and resolution on homologous series of alkylagaroses.
Implantation of hydrocarbon chains on beaded agarose results in column materials capable of adsorbing erythrocytes or lymphocytes from various sources. In a homologous series of such columns, identical in all structural respects (ligand density, charge density and ultrastructure) except for the length of their hydrocarbon chains, the capacity to adsorb these cells generally increases with increase in the number of carbon atoms per chain. The cells can be desorbed from the columns by repeated gentle pipetting in the presence of bovine serum albumin (for erythrocytes) or foetal calf serum (for lymphocytes). Under the conditions used for adsorption and desorption there is neither physical entrapment of the cells in the column nor apparent damage to their integrity, as indicated by the facts that (a) 95-98% of the cells applied on the column can be recovered; (b) erythrocytes eluted from the column and those yet to be applied are morphologically indistinguishable, exhibit an identical osmotic fragility profile and, (after desorption) retain the same adsorption profile for the columns; and (c) over 95% of the mouse spleen lymphocytes, eluted from the columns, continue to exclude the dye Trypan blue, suggesting that they preserve their viability. Cells from different sources exhibit different adsorption profiles on homologous series of alkylagaroses, allowing the detection of differences in the surface of these cells and their resolution by these columns. Exploratory experiments with artificial cell mixtures are described, illustrating the possibility of enriching the mixture with one of the cell types by preferential adsorption or exclusion on an appropriate alkylagarose. The best resolution was obtained with a mixture of erythrocytes and spleen lymphocytes (both from DBA/1 mice). After appropriate "tailoring" of the column material used (an ethylagarose) it was possible to apply a 1:1 mixture of these cells and to obtain (after preferential adsorption) essentially pure erythrocytes (98% of the excluded cells).(ABSTRACT TRUNCATED AT 400 WORDS)
D000327:Adsorption; D000818:Animals; D002449:Cell Aggregation / Q000187:drug effects; D002469:Cell Separation; D002477:Cells / Q000032:analysis*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002645:Chickens; D002845:Chromatography; D003488:Cyanogen Bromide; D003584:Cytological Techniques; D004910:Erythrocyte Membrane / Q000032:analysis; D006168:Guinea Pigs; D066298:In Vitro Techniques; D008214:Lymphocytes / Q000032:analysis; D051379:Mice; D008811:Mice, Inbred DBA; D019158:N-Acetylneuraminic Acid; D009439:Neuraminidase; D009996:Osmotic Fragility; D011107:Polylysine; D011817:Rabbits; D012685:Sepharose; D012794:Sialic Acids / Q000097:blood; D013154:Spleen / Q000166:cytology
D012794:Sialic Acids; D011107:Polylysine; D012685:Sepharose; D009439:Neuraminidase; D019158:N-Acetylneuraminic Acid; D003488:Cyanogen Bromide