Synthesis of steroid tracers with a oxyme ligation method and use thereof in fluorescent polarisation immunoassay

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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.

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About the authors

I. A. Prokhorenko

Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry RAS

Author for correspondence.
Email: prig67@mail.ru
Russian Federation, ul. Miklukho-Maklaya 16/10, Moscow, 117997

D. A. Glushchenko

Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry RAS

Email: prig67@mail.ru
Russian Federation, ul. Miklukho-Maklaya 16/10, Moscow, 117997

E. L. Gulyak

Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry RAS

Email: prig67@mail.ru
Russian Federation, ul. Miklukho-Maklaya 16/10, Moscow, 117997

I. V. Mikhura

Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry RAS

Email: prig67@mail.ru
Russian Federation, ul. Miklukho-Maklaya 16/10, Moscow, 117997

V. A. Korshun

Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry RAS

Email: prig67@mail.ru
Russian Federation, ul. Miklukho-Maklaya 16/10, Moscow, 117997

L. I. Mukhametova

Lomonosov Moscow State University

Email: prig67@mail.ru

Department of Chemistry

Russian Federation, Leninskiye Gory 1/3, Moscow, 119991

S. А. Eremin

Lomonosov Moscow State University

Email: prig67@mail.ru

Department of Chemistry

Russian Federation, Leninskiye Gory 1/3, Moscow, 119991

References

  1. Zamri M.F.M.A., Bahru R., Pramanik S.K., Fattah I.M.R. // J. Water Proc. Engin. 2021. V. 41. 102017. https://doi.org/10.1016/j.jwpe.2021.102017
  2. Miller W.L., Auchus R.J. // Endocr. Rev. 2011. V. 32. P. 81. https://doi.org/10.1210/ER.2010-0013
  3. Куджаев А.М., Андрианова А.Г., Гущина А.Е., Смирнов И.В., Ротанова Т.В. // Биоорг. химия. Т. 48. С. 381–414. [Kudzhaev A.M., Andrianova A.G., Gustchina A.E., Smirnov I.V., Rotanova T.V. // Russ. J. Bioorg. Chem. 2022. V. 48. P. 678–709.] https://doi.org/10.1134/S1068162022040136
  4. Gore A.C. // JAMA Int. Med. 2016. V. 176. P. 1705–1706. https://doi.org/10.1001/jamainternmed.2016.5766
  5. Abraham G.E. // J. Steroid Biochem. 1975. V. 6. P. 261–270. https://doi.org/10.1016/0022-4731(75)90141-7
  6. Relave F., Lefebvre R.C., Beaudoin S., Price C. // Can. Vet. J. 2007. V. 48. P. 823–826.
  7. Stanczyk F.Z., Lee J.S., Santen R.J. // Cancer Epidemiol. Biomarkers Prev. 2007. V. 16. P. 1713–1719. https://doi.org/10.1158/1055-9965.EPI-06-0765
  8. Tate J., Ward G. // Clin. Biochem. Rev. 2004. V. 25. P. 105–120.
  9. Hall M.D., Yasgar A., Peryea T., Braisted J.C., Jadhav A., Simeonov A., Coussens N.P. // Methods Appl. Fluoresc. 2016. V. 4. P. 022001. https://doi.org/10.1088/2050-6120/4/2/022001
  10. Smith D.S., Eremin S.A. // Anal. Bioanal. Chem. 2008. V. 391. P. 1499–1507. https://doi.org/10.1007/s00216-008-1897-z
  11. Choi M.J., Choi J., Yoon D.Y., Park J., Eremin S.A. // Biol. Pharm. Bull. 1997. V. 20. P. 309–314. https://doi.org/10.1248/bpb.20.309
  12. Колосова A.Ю., Еремин С.А., Гаврилова Е.М., Егоров А.М. // Проблемы эндокринологии. 1994. Т. 40. С. 48–51. https://doi.org/10.14341/probl12147
  13. Hong J.Y., Choi M.J. // Biol. Pharm. Bull. 2002. V. 25. P. 1258–1262. https://doi.org/10.1248/bpb.25.1258
  14. Quémener D., Davis T.P., Barner-Kowollik C., Stenzel M.H. // Chem. Commun. 2006. V. 48. P. 5051–5053. https://doi.org/10.1039/B611224B
  15. Clède S., Lambert F., Sandt C., Kascakova S., Unger M., Harté E., Plamont M.-A., Saint-Forth R., Deniset- Besseau A., Guerouiab Z., Hirschmugl C., Lecomte S., Dazzi A., Vessières A., Policar C. // Analyst. 2013. V. 138. P. 5627–5638. https://doi.org/10.1039/C3AN00807J
  16. Robinson S.W., Beer P.D. // Org. Biomol. Chem. 2017. V. 15. P. 153–159. https://doi.org/10.1039/ c6ob02339h
  17. Khomutov A.R., Vepsalainen J.J., Shvetsov A.S., Hyvönen T., Keinänen T.A., Pustobaev V.N., Eloranta T.O., Khomutov R.M. // Tetrahedron. 1996. V. 52. P. 13751–13766. https://doi.org/10.1016/0040-4020(96)00836-8
  18. Sapozhnikova A., Misyurin V.A., Ryazantsev D.Y., Kokin E.A., Finashutina Yu.P., Alexeeva A.V., Ivanov I.A., Kocharovskaya M.V., Tikhonova N.A., Popova G.P., Alferova V.A., Ustinov A.V., Korshun V.A., Brylev V.A. // Int. J. Mol. Sci. 2022. V. 22. P. 12845. https://doi.org/10.3390/ijms222312845
  19. Kolmer D.K., Cool E.T. // Chem. Rev. 2017. V. 117. P. 10358–10376. https://doi.org/10.1021/acs.chemrev.7b00090
  20. Adamczyk M., Chen Y.-Y., Grote J., Mattingly P.G. // Steroids. 1999. V. 64. P. 283–290. https://doi.org/10.1016/S0039-128X(99)00004-5
  21. Мухаметова Л.И., Крылов В.Б., Соловьев A.C., Яшунский Д.В., Матвеев A.Л., Тикунова Н.В., Еремин С.А., Нифантьев Н.Э. // Известия Акад. наук. Сер. хим. 2021. №. 5. С. 975–981.
  22. Zhao Q., Bai Y., Wang H. // Talanta. 2020. V. 217. P. 121018. https://doi.org/10.1016/j.talanta.2020.121018

Supplementary files

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1. JATS XML
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2. 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.

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3. Fig. 2. Stereoisomeric fragments of oximes (VII) and (VIII).

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4. 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 ).

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5. 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.

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6. 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.

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7. Scheme 1. Synthesis of bifunctional linker reagent (V).

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8. Scheme 2. Synthesis of the ethoxyethylidene-protected aminooxy derivative (VI) of 6-carboxyfluorescein (6-FAM) and fluorescent tracers (VII) and (VIII).

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