A Sandwich Enzyme-Linked Immunoabsorbent Assay for Measurement of Gonadotropin-Releasing Hormone-Toxin Conjugates
2006
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 <20 min).
Conclusion The assays were found to be specific, sensitive and accurate for measurement of GnRH-toxin conjugates in vitro and in vivo.
Keywords:
- Correction
- Source
- Cite
- Save
- Machine Reading By IdeaReader
30
References
1
Citations
NaN
KQI