1. A Sandwich Enzyme-Linked Immunoabsorbent Assay for Measurement of Gonadotropin-Releasing Hormone-Toxin Conjugates
- Author
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Matthew C. Allen, L. Michael Glode, Terry M. Nett, Maciej Wieczorek, and Wei-Hsiung Yang
- Subjects
endocrine system ,Time Factors ,RNase P ,Immunology ,Antiviral protein ,Enzyme-Linked Immunosorbent Assay ,Gonadotropin-releasing hormone ,Biology ,Sensitivity and Specificity ,Gonadotropin-Releasing Hormone ,In vivo ,Animals ,Immunology and Allergy ,Cytotoxicity ,chemistry.chemical_classification ,Sheep ,Cytotoxins ,Obstetrics and Gynecology ,Molecular biology ,In vitro ,Enzyme ,Reproductive Medicine ,chemistry ,Rabbits ,hormones, hormone substitutes, and hormone antagonists ,Conjugate - Abstract
Problem Biological effectiveness of targeted cytotoxins is dependent on their stability, circulating half-life, receptor binding ability, and cytotoxicity. The objective of this study was to compare stability of gonadotropin-releasing hormone (GnRH)-toxin conjugates made with disulfide linkers to those using a maleimidodibutyryl (mb) linkage. Method of study We developed a sandwich enzyme-linked immunoabsorbent assay recognizing both GnRH analog and cytotoxin to ensure the conjugate measured was intact. Anti-D-Leu6-GnRH was used for capture and anti-pokeweed antiviral protein (anti-PAP) or anti-RNase for quantification. Specificity was verified by lack of reactivity with ovine FSH and LH, PAP, RNase, and D-Lys6-GnRH. Results Conjugates prepared using disulfide linkages were not stable in serum in vitro (half-lives 2 hr. Clearance of mbGnRH-PAP from the circulation of sheep was rapid (t1/2 more...
- Published
- 2006
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