Submitted to: Journal of Agricultural and Food Chemistry
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/25/2005
Publication Date: 5/18/2005
Citation: Shelver, W.L., Keum, Y., Kim, H., Rutherford, D., Hakk, H., Bergman, A., Li, Q.X. 2005. Hapten syntheses and antibody generation for the development of a polybrominated flame retardant elisa. Journal of Agricultural and Food Chemistry 53(10):3840-3847.
Interpretive Summary: This work describes the use of computer modeling to develop antibodies to measure the amount of flame retardant in various samples. The antibodies form the basis of a rapid and specific analysis (ELISA) that could be used to rapidly screen environmental and food samples. This rapid screening is necessary to run the large number of samples necessary to determine if these compounds are accumulating in our environment or contaminating our food samples. The computer modeling was tested for its ability to predict the antibodies performance and speed the development of the assay. The test utilized seven different antibodies generated from four compounds in goats and rabbits. The computer modeling correctly predicted most of the antibody properties and can be used to assist scientists in development of new antibody-based assays.
Technical Abstract: Polybrominated diphenyl ethers (PBDEs) are a class of brominated flame retardants that are of increasing environmental concern. Several antibodies were developed for the polybrominated diphenyl ether flame retardant BDE-47 (1), often found in the highest concentration in human milk, plasma, and adipose tissue. Four haptens with different bromine and linker substitution patterns were synthesized and utilized to generate five polyclonal antibodies from goats and two polyclonal antibodies from rabbits. Competition was assessed using four different coating antigens for all seven antibodies. The coating antigen showed marked effects on competition, generally when the same hapten was used for antibody and the coating antigen less competition was observed. The effect of BDE structure on competition was evaluated by using BDE-47 (1), BDE-99 (2), BDE-100 (3), BDE-153 (4), and BDE-183 (5). None of the compounds showed high competition with antibody I-KLH, presumably because steric hindrance prevented formation of an efficient binding site. As predicted from structural considerations, BDE-47 (1) competed well with the remaining antibodies, whereas BDE-100 (3) competed well with only II-KLH. The remaining congeners (BDE-99 (2), BDE-153 (4), and BDE-183 (5)) contain bromines that cannot be positioned in binding sites and thus cross react poorly. The competition study demonstrated that a bromine substitution on the congener could occupy a position analogous to the linker's position.