Preparation of Dinitrophenyl- and Trinitrophenyl-Labeled Erythrocytes

Abstract

The introduction of new immuno-determinant groups, such as dinitrophenyl (DNP) and trinitrophenyl (TNP), into whole cells became possible through the use of the DNP and TNP sulfonic acids. The chemical reaction through which a covalent linkage is established between the protein and the DNP or TNP hapten is a relatively mild nucleophilic substitution. Electron donating functional groups, in the carrier molecule, such as SH^- or NH<Stack><Subscript>2</Subscript><Superscript>-</Superscript></Stack> or OH^- groups will react with electron accepting sulfonic acid radicals on the nitrated benzene ring, thus forming a stable covalent bond between the hapten and the carrier. DNP and TNP sulfonic acids are soluble in water or saline and the reaction will proceed smoothly at room temperature or at slightly elevated temperatures. The treatment of a carrier molecule or cell with these reagents will not introduce considerable unwanted changes in their structure.