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.
| Identifer | oai:union.ndltd.org:UMASS/oai:scholarworks.umass.edu:dissertations-8523 |
| Date | 01 January 1992 |
| Creators | Mei, Joanne Virginia |
| Publisher | ScholarWorks@UMass Amherst |
| Source Sets | University of Massachusetts, Amherst |
| Language | English |
| Detected Language | English |
| Type | text |
| Source | Doctoral Dissertations Available from Proquest |
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