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Immunochemical detection of flavonoid glycosides: development, specificity, and application of novel monoclonal antibodies.
- Source :
-
Archives of biochemistry and biophysics [Arch Biochem Biophys] 2008 Aug 15; Vol. 476 (2), pp. 124-32. Date of Electronic Publication: 2008 Feb 05. - Publication Year :
- 2008
-
Abstract
- Flavonoid-rich diets are expected to decrease the risk of cardiovascular diseases. The localization and target sites of flavonoids underlying the protective mechanism in vivo have not been fully investigated because the methods for detection of flavonoids have been limited to chemical analysis such as high-performance liquid chromatography. To further understand the actions of flavonoids in vivo, we developed a novel methodology that immunochemically evaluates flavonoids using specific antibodies. Quercetin-3-glucuronide (Q3GA), a major metabolite in human plasma, was coupled with keyhole limpet hemocyanin. Alternatively, the sugar moiety of quercetin-3-glucoside (Q3G) was succinylated and then coupled with a carrier protein. Using these two immunogens, we finally obtained two monoclonal antibodies, mAb14A2 and mAb11G6, from the immunogen using Q3GA and Q3G, respectively. Competitive enzyme-linked immunosorbent assay showed the unique difference in the specificity between the two similar antibodies: mAb14A2 recognized several quercetin-3-glycosides including Q3G and rutin but mAb11G6 was highly specific to the Q3G structure. The macrophage-derived foam cells in human atherosclerotic lesions were significantly stained with mAb14A2 but scarcely with mAb11G6. These results showed that the anti-flavonoid glycoside antibodies are useful tools for evaluating their localization in tissues and that the specificities strongly depend on the immunogen design for synthesizing the hapten-protein conjugates.
- Subjects :
- Animals
Atherosclerosis blood
Atherosclerosis pathology
Enzyme-Linked Immunosorbent Assay
Foam Cells metabolism
Foam Cells pathology
Glucuronides blood
Glucuronides pharmacology
Hemocyanins metabolism
Humans
Macrophages pathology
Mollusca
Quercetin blood
Quercetin metabolism
Sensitivity and Specificity
Antibodies, Monoclonal chemistry
Flavonoids immunology
Glycosides immunology
Subjects
Details
- Language :
- English
- ISSN :
- 1096-0384
- Volume :
- 476
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Archives of biochemistry and biophysics
- Publication Type :
- Academic Journal
- Accession number :
- 18267102
- Full Text :
- https://doi.org/10.1016/j.abb.2008.01.022