Succinylation of hapten-protein conjugates facilitates coupling to erythrocytes by water soluble carbodiimide: Preparation of stable and sensitive target cells for use in hemolytic assays☆

Abstract A facile method is described for the preparation of haptenated sheep red blood cells (SRBC) for use as targets in hemolytic spot and plaque assays for the detection of antihapten antibody. The method involves the use of the water soluble 1-ethyl-3-(3-dimethyl-aminopropyl)carbodiimide (EDCI) as a reagent to couple hapten-succinyl-rabbit serum albumin conjugates to SRBC. The presence of the succinyl groups on such conjugates is shown to increase the efficacy of the resulting target cells, presumably by acting as a substrate for the EDCI and thus increasing the extent of coupling to SRBC.