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.
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 anti-hapten 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.
D000818:Animals; D000906:Antibodies; D002234:Carbodiimides / Q000276:immunology*; D002470:Cell Survival; D004912:Erythrocytes / Q000276:immunology*; D006241:Haptens; D006461:Hemolysis; D006462:Hemolytic Plaque Technique; D006822:Hybrid Cells / Q000276:immunology; D011817:Rabbits; D012709:Serum Albumin / Q000276:immunology; D012756:Sheep; D012995:Solubility; D013387:Succinic Anhydrides / Q000494:pharmacology
D000906:Antibodies; D002234:Carbodiimides; D006241:Haptens; D012709:Serum Albumin; D013387:Succinic Anhydrides