Synthesis of steroid tracers with a oxyme ligation method and use thereof in fluorescent polarisation immunoassay
| Dublin Core | PKP Metadata Items | Metadata for this Document | |
| 1. | Title | Title of document | Synthesis of steroid tracers with a oxyme ligation method and use thereof in fluorescent polarisation immunoassay |
| 2. | Creator | Author's name, affiliation, country | I. A. Prokhorenko; Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry RAS; Russian Federation |
| 2. | Creator | Author's name, affiliation, country | D. A. Glushchenko; Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry RAS; Russian Federation |
| 2. | Creator | Author's name, affiliation, country | E. L. Gulyak; Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry RAS; Russian Federation |
| 2. | Creator | Author's name, affiliation, country | I. V. Mikhura; Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry RAS; Russian Federation |
| 2. | Creator | Author's name, affiliation, country | V. A. Korshun; Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry RAS; Russian Federation |
| 2. | Creator | Author's name, affiliation, country | L. I. Mukhametova; Lomonosov Moscow State University; Russian Federation |
| 2. | Creator | Author's name, affiliation, country | S. А. Eremin; Lomonosov Moscow State University; Russian Federation |
| 3. | Subject | Discipline(s) | |
| 3. | Subject | Keyword(s) | antibodies; steroid hormones; homogeneous immunoassay; fluorescent dyes; oxime ligation |
| 4. | Description | Abstract | A method for the synthesis of ketosteroid oxime conjugates with a fluorescein (6-FAM) derivative connected via a short bifunctional aminooxypropyl linker is proposed. The conjugates were used as tracers for fluorescence polarization immunoassay (FPIA) with monoclonal antibodies. Stereoisomers by the oxime group were separated by preparative RP TLC and HPLC and tested in the FPIA method. Binding parameters of of tracers with monoclonal antibodies (mAbs) to progesterone were studied. Tracers showed high efficiency in the analysis of steroid hormones, the detection limit of progesterone being lower than that for a previously described analog. The Z-isomer was found to be more sensitive in the FPIA analysis compared to the E-isomer. |
| 5. | Publisher | Organizing agency, location | The Russian Academy of Sciences |
| 6. | Contributor | Sponsor(s) | |
| 7. | Date | (DD-MM-YYYY) | 19.08.2024 |
| 8. | Type | Status & genre | Peer-reviewed Article |
| 8. | Type | Type | Research Article |
| 9. | Format | File format | |
| 10. | Identifier | Uniform Resource Identifier | https://rjeid.com/0132-3423/article/view/671002 |
| 10. | Identifier | Digital Object Identifier (DOI) | 10.31857/S0132342324010055 |
| 10. | Identifier | eLIBRARY Document Number (EDN) | OWPZNS |
| 11. | Source | Title; vol., no. (year) | Bioorganičeskaâ himiâ; Vol 50, No 1 (2024) |
| 12. | Language | English=en | ru |
| 13. | Relation | Supp. Files |
Fig. 1. Structure of previously described tracers for FPIA based on steroid hormones: (a) – Pg3CMO-EDF (progesterone carboxymethyloxime amide with ethylenediamine fluorescein thiocarbamate); (b) – 11-α-hydroxyprogesterone, acylated with succinic anhydride. (137KB) Fig. 2. Stereoisomeric fragments of oximes (VII) and (VIII). (58KB) Fig. 3. Change in the fluorescence polarization signal of free tracers E(VII), Z(VII) and Pg3CMO-EDF (2.5 nM) during the formation of complexes with specific antibodies to progesterone MAb-Pg (20 nM) in comparison with nonspecific MAb-3G11 (30 nM ). (197KB) Fig. 4. Dependence of changes in the fluorescence polarization signal on the concentration of antibodies for tracers E(VII) (curve 1), Z(VII) (curve 2) and Pg3CMO-EDF (curve 3) (tracer concentration 2.5 nM); pH 8.5, 25°C. (198KB) Fig. 5. Calibration dependences (a) and normalized calibration dependences (b) for tracers E(VII) (curve 1), Z(VII) (curve 2) and Pg3CMO-EDF (curve 3) (tracer concentrations 2.5 nM); pH 8.5, 25°C. (332KB) Scheme 1. Synthesis of bifunctional linker reagent (V). (111KB) Scheme 2. Synthesis of the ethoxyethylidene-protected aminooxy derivative (VI) of 6-carboxyfluorescein (6-FAM) and fluorescent tracers (VII) and (VIII). (205KB) |
| 14. | Coverage | Geo-spatial location, chronological period, research sample (gender, age, etc.) | |
| 15. | Rights | Copyright and permissions |
Copyright (c) 2024 Russian Academy of Sciences |