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Immunoassays for des-Arg9-bradykinin
C E Odya1, R J Carlin, R D Yapa
1Medical Sciences Program, Indiana University School of Medicine, Bloomington 47405.
Journal of Immunoassay
|December 1, 1993
Summary
Researchers developed a highly sensitive radioimmunoassay for detecting des-Arg9-bradykinin. This assay, using specific monoclonal antibodies, offers improved detection of bradykinin metabolites.
Area of Science:
- Immunology
- Biochemistry
- Assay Development
Background:
- Bradykinin is a vasoactive peptide involved in various physiological processes.
- Measuring bradykinin metabolites like des-Arg9-bradykinin is crucial for understanding kinin system activity.
- Existing methods for detecting bradykinin metabolites may lack sensitivity or specificity.
Purpose of the Study:
- To develop and characterize monoclonal antibodies for detecting des-Arg9-bradykinin.
- To establish a highly sensitive radioimmunoassay for quantifying des-Arg9-bradykinin.
- To assess the specificity of the developed antibodies and assay.
Main Methods:
- Hybridoma technology was used to generate monoclonal antibodies against des-Arg9-bradykinin.
- Enzyme-linked immunosorbent assays (ELISAs) were employed for initial screening and affinity determination.
- Radioimmunoassays (RIAs) were developed using labeled antigens and dextran-coated charcoal separation.
Main Results:
- Seventy-nine hybridomas producing kinin-reactive antibodies were identified.
- All antibodies showed higher reactivity towards des-Arg9-bradykinin than bradykinin.
- A highly sensitive RIA (0.08 +/- 0.03 nM IC50) was developed using clone OLNBK-5, with excellent specificity for des-Arg9-bradykinin (100%) over bradykinin (29%).
Conclusions:
- Monoclonal antibody clone OLNBK-5 is highly specific and sensitive for des-Arg9-bradykinin.
- The developed radioimmunoassay provides a robust tool for quantifying des-Arg9-bradykinin.
- This assay has potential applications in research involving the kinin system.