The production of antibodies to a veterinary drug requires conjugation of the low molecular weight compound to a larger molecular weight substance (carrier protein) to enable immune recognition. The resulting immunogen is usually mixed with an adjuvant to create a prepnration thnt will elicit a strong and lasting inullune response to the complex when administered to the host animal. Fretmd's adjuvant has been previously used but can produce adverse side effects in the host. A replacement adjuvant for Fretllld's was sought: Montanide ISA 50V was found to be suitable for the production of antibodies to three veterinary clmg haptens without the adverse effects associated with Freund·s. Antibodies were required for the development of inummoassays for the coccidiostats diclazuril and . robenidine. TIle lack of a suitable fimctional group for coupling to the protein made immtlllogen preparation difficult for both these compounds. lvlimics possessing part of the relevant structures and more open to chemical manipulation were used as haptens to prepare inummogens and subsequently specific antibodies for diclazuril and robenidine were obtained. Polyclonal antisera to tlle antibiotic chloramphenicol were successfully raised in three different species. All of the antisera were affected to a similar e;-,.1ent when sanlple matrix was introduced to the assays. It was found that a heterologous format of both the ELISA and biosensor assay increased tlle crossreactivity of donkey and goat antibodies but not emile!. A camel antibody was separated into its three IgG subclasses. The conventional four chain molecule displayed affinity to chloramphenicol and the carrier protein \\rule the two heavy chain IgGs were only able to bind the carrier. TIle use of a suitable adjunult. antigen mimics and appropriate chemical manipulation techniques can enable the researcher to generate specific and sensitive antibodies to a range of small molecular weight compounds.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:491939 |
| Date | January 2008 |
| Creators | Fodey, Terence Lee |
| Publisher | Queen's University Belfast |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
Page generated in 0.0092 seconds