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The use of enzyme immunoassay for the quantitative analysis of an insect growth regulator and insect juvenile hormone from environmental and biological matrices
Abstract
Two immunoassay formats were developed for the detection of low levels of the insect growth regulator, methoprene. The generation of methoprene-specific antibodies needed for such assays relied on the preparation of a methoprene-carrier immunogen. 11-Methoxy-3,7,11-trimethyl-2E,4E-dodecadienoic acid was covalently bound to a protein carrier via a spacer group. Two activated ester methods were used to prepare the immunogen, one of which forms a water soluble, activated ester of methoprene. Polyclonal antibodies raised against the methoprene immunogen were highly specific for methoprene. An indirect enzyme-linked immunosorbent assay (iELISA) and a competitive inhibition enzyme immunoassay (CIEIA) were developed using the polyclonal antisera. The range of the methoprene indirect ELISA was from 5 to 300 ng/mL (ppb), with an I$\sb{50}$ of 50 ng/mL, while the CIEIA has a range from 1.0 to 10 ppb, with an I$\sb{50}$ of 3.5 ppb. An indirect ELISA was also developed for insect juvenile hormone III from rabbit polyclonal antisera. The synthesis of several juvenile hormone derivatives used to prepare a juvenile hormone immunogen is described. The immunogen consisted of juvenile hormone III bound to a spacer arm via an ester, which was bound, in turn to a carrier protein via an amide bond. The resulting immunochemical assay showed high specificity for juvenile hormone III, with an I$\sb{50}$ of 225 ng/well. The juvenile hormone homologs, I and II, had I$\sb{50}$'s of 5000 and 800 ng/well, respectively. Other juvenile hormone analogs cross reacted with the juvenile hormone III antibody to a much lesser degree. The juvenile hormone III indirect ELISA has great potential for becoming the first readily available, easy-to-use analytical technique for the quantification of the hormone from biological materials.
Subject Area
Entomology|Environmental science|Chemical engineering
Recommended Citation
Mei, Joanne Virginia, "The use of enzyme immunoassay for the quantitative analysis of an insect growth regulator and insect juvenile hormone from environmental and biological matrices" (1992). Doctoral Dissertations Available from Proquest. AAI9219467.
https://scholarworks.umass.edu/dissertations/AAI9219467