Cell affinity chromatography with ligands immobilized through cleavable mercury-sulfur bonds.
A new methodology for cell separation by affinity chromatography is described. We have conjugated the organomercurial mersalyl to trisacryl beads bearing primary amino groups. Thiolated ligands can be immobilized on this matrix through cleavable Hg-S bonds. Two model studies of cell separation are reported: (i) concanavalin A thiolated with N-succinimidyl-3-(2-pyridyldithio)-propionate and immobilized on mersalyl-trisacryl; mouse thymocytes bound to Con A-mersalyl-trisacryl were eluted from the support by short thiol treatment which preserved cell viability; (ii) anti-dinitrophenyl antibodies modified with S-acetyl-mercaptosuccinic anhydride and immobilized on mersalyl-trisacryl; sheep erythrocytes, previously labelled with trinitrobenzene sulfonic acid, bound to this support and were easily recovered by thiol treatment without hemolysis. This methodology should overcome difficulties frequently encountered in cell affinity chromatography. Cell support multivalent interactions or high affinity of cell-ligand bindings often require drastic elution conditions which prevent viable cell recovery.
D000818:Animals; D002469:Cell Separation / Q000379:methods*; D002846:Chromatography, Affinity / Q000379:methods*; D003208:Concanavalin A; D004912:Erythrocytes; D008297:Male; D008628:Mercury; D008634:Mersalyl; D051379:Mice; D009941:Organomercury Compounds; D012756:Sheep; D013438:Sulfhydryl Compounds; D013601:T-Lymphocytes
D009941:Organomercury Compounds; D013438:Sulfhydryl Compounds; D003208:Concanavalin A; D008634:Mersalyl; D008628:Mercury