Abstract
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.
Full text
PDF
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ballmer-Hofer K., Schlup V., Burn P., Burger M. M. Isolation of in situ crosslinked ligand-receptor complexes using an anticrosslinker specific antibody. Anal Biochem. 1982 Nov 1;126(2):246–250. doi: 10.1016/0003-2697(82)90511-5. [DOI] [PubMed] [Google Scholar]
- Bauminger S., Wilchek M. The use of carbodiimides in the preparation of immunizing conjugates. Methods Enzymol. 1980;70(A):151–159. doi: 10.1016/s0076-6879(80)70046-0. [DOI] [PubMed] [Google Scholar]
- Boraker D. K., Stinebring W. R., Kunkel J. R. BrucELISA: an enzyme-antibody immunoassay for detection of Brucella abortus antibodies in milk: correlation with the Brucella ring test and with shedding of viable organisms. J Clin Microbiol. 1981 Oct;14(4):396–403. doi: 10.1128/jcm.14.4.396-403.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- GOODFRIEND T. L., LEVINE L., FASMAN G. D. ANTIBODIES TO BRADYKININ AND ANGIOTENSIN: A USE OF CARBODIIMIDES IN IMMUNOLOGY. Science. 1964 Jun 12;144(3624):1344–1346. doi: 10.1126/science.144.3624.1344. [DOI] [PubMed] [Google Scholar]
- König W., Geiger R. Eine neue Methode zur Synthese von Peptiden: Aktivierung der Carboxylgruppe mit Dicyclohexycarbodiimid unter Zusatz von 1-Hydroxy-benzotriazolen. Chem Ber. 1970;103(3):788–798. doi: 10.1002/cber.19701030319. [DOI] [PubMed] [Google Scholar]
- Lizana J., Hellsing K. Polymer enhancement of automated immunological nephelometric analysis, as illustrated by determination of urinary albumin. Clin Chem. 1974 Apr;20(4):415–420. [PubMed] [Google Scholar]
- Paul D., Flouret G., Tomita J. T., Ranney K., Schenck J., Anderson B. Preparation and specificities of antisera to the amino-terminal sequence of the carcinoembryonic antigen. Cancer Res. 1981 Jun;41(6):2315–2321. [PubMed] [Google Scholar]
- Timkovich R. Detection of the stable addition of carbodiimide to proteins. Anal Biochem. 1977 May 1;79(1-2):135–143. doi: 10.1016/0003-2697(77)90387-6. [DOI] [PubMed] [Google Scholar]
- Wofsy L. Methods and applications of hapten-sandwich labeling. Methods Enzymol. 1983;92:472–488. doi: 10.1016/0076-6879(83)92040-2. [DOI] [PubMed] [Google Scholar]
- Wojtkowiak Z., Briggs R. C., Hnilica L. S. A sensitive method for staining proteins transferred to nitrocellulose sheets. Anal Biochem. 1983 Mar;129(2):486–489. doi: 10.1016/0003-2697(83)90581-x. [DOI] [PubMed] [Google Scholar]