Antisera specificities to 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide adducts of proteins.
Animals were immunized with either 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI)-coupled peptide hapten-protein carrier conjugates or with N-acylurea EDCI derived protein carrier. Sera from all animals demonstrated antibody to N-acylurea EDCI when tested by double immunodiffusion using N-acylurea EDCI coupled to a heterologous carrier. Two rabbit antisera, one produced to EDCI-coupled peptide-24-thyroglobulin and another produced to N-acylurea EDCI derived thyroglobulin were further characterized using an enzyme-linked immunosorbent assay (ELISA). Competitive binding assays using either EDCI-coupled peptide-protein conjugates or N-acylurea EDCI derived proteins demonstrated the presence of N-acylurea EDCI reactive antibodies. Such antibodies were also shown to react with the EDCI isourea employed as inhibitor. The specificity of the anti-EDCI antibodies to portions of the EDCI molecule was demonstrated using structural analogues of EDCI. Both 3-dimethylaminopropylamine and 2-dimethylaminoethanol effectively inhibited anti-N-acylurea EDCI reactivity. These results show that when EDCI is used as a coupling agent, antibody is produced to N-acylurea EDCI-carrier, the antibody being primarily directed to the 3-dimethylaminopropyl end of the EDCI molecule.
D000818:Animals; D000917:Antibody Formation; D000918:Antibody Specificity; D001667:Binding, Competitive; D002234:Carbodiimides / Q000276:immunology*; D002352:Carrier Proteins / Q000276:immunology*; D005022:Ethyldimethylaminopropyl Carbodiimide / Q000031:analogs & derivatives / Q000276:immunology*; D006241:Haptens / Q000276:immunology*; D007106:Immune Sera / Q000276:immunology*; D011817:Rabbits; D013329:Structure-Activity Relationship
D002234:Carbodiimides; D002352:Carrier Proteins; D006241:Haptens; D007106:Immune Sera; D005022:Ethyldimethylaminopropyl Carbodiimide