Isolation of dinitrophenyl-skin protein conjugates by immunoadsorbent chromatography.

Immunoadsorbent columns charged with rabbit anti-dinitrophenyl (DNP) antibody were used to isolate and concentrate DNP-protein conjugates from the 105,000 g supernate of extracts prepared from guinea pig epidermis previously painted with dinitrofluorobenzene (DNFB). Conjugates isolated in this manner elicited positive delayed hypersensitivity skin tests when injected intradermally into DNFB-sensitized animals. Dinitrophenylated epidermal protein conjugates prepared in vitro were capable of specifically inhibiting the migration of peritoneal exudate cells obtained from DNFB-sensitized guinea pigs. These results demonstrate the application of immunoadsorbent chromatography and the migration inhibition assay to the further definition of the epitopes involved in contact sensitivity.