Polarization fluoroimmunoassays of progesterone
https://doi.org/10.14341/probl12147
Abstract
A quick reliable homogenous polarization fluoroimmunoassay (PFIA) of progesterone was developed. The assay is carried out with Abbott TDx (USA) polarization fluorometer and it takes 5-7 min to analyze 10 samples by this method. The range of progesterone concentrations determined is 1 to 1000 ng/ml. Fluorescein labeled progesterone-3-carbo- xymethyloxime which was used as tracer (labeled antigen) during analysis has been synthesized and purified. Two types of PFIA were developed: one making use of rabbit antiserum to progesterone-3-carboxymethyloxime conjugated with bovine serum albumin (BSA) or keyhole limpet hemocianin (KLH), in the other antiserum to progesterone-11-hemisuccinate-BSA (or KLH) is used. Different combinations of the tracer and antibodies were used. The sensitivity of heterologous PFIA (with antibodies to immunogen heteroioguos to tracer by structure) was higher than that of homologous PFIA. The test is sufficienctly sensitive and specific. The method is particularly valuable for determination of progesterone in model solutions (using buffer standards).
About the Authors
A. Yu. Kolosova
Lomonosov Moscow State University
Russian Federation
Russian Federation
S. A. Yeremin
Lomonosov Moscow State University
Russian Federation
Russian Federation
Ye. M. Gavrilova
Lomonosov Moscow State University
Russian Federation
Russian Federation
A. M. Yegorov
Lomonosov Moscow State University
Russian Federation
Russian Federation
References
1. Бекбергенов Б. М., Житников В. Т. // Антибиотики и химиотер,— 1988,— Т. 33, № 1,— С. 72—76.
2. Новые методы иммуноанализа / Тертон М., Бангхем Д. Р., Колкотт К. А. и др.; Пер. с англ.— М., 1991.
3. Теория и практика иммуноферментного анализа / Егоров А. М., Осипов А. П., Дзантиев Б. Б., Гаврилова Е. М.— М„ 1991.
4. Bourque J., Sulon J., Demey-Ponsart E. et al. // Clin. Chem.— 1986,—Vol. 32, N 6.— P. 948—951.
5. Dandliker W. B., Schapiro H. C„ Meduski J. W. et al. // Immunochemistry.— 1964.— Vol. 1.— P. 165—191.
6. Dray F., Andrieu J.-M., Renaud F. // Biochim. biophys. Acta.— 1975,—Vol. 403,— P. 131 — 136.
7. Joice B. J., Gracham F. R., Farmey D. R. // Steroids.— 1977,—Vol. 29, N 5.— P. 761—767.
8. Methods of Enzymatic Analysis.— 3-d Ed / Ed. H. U. Berg- meyer.— Weinheim, 1985.— Vol. 8.— P. 212—222.
9. Mitsuma M., Kambegawa A., Okinaga S., Arai K. // J. Steroid Biochem.— 1987.— Vol. 28, N 1.— P. 83—88.
10. Munro C., Stabenfeldt G. // J. Endocr.— 1984.— Vol. 101.— P. 41—49.
11. Pourfarzaneh M., White G. W., Landon J., Smith D. S. 11 Clin. Chem.— 1980.— Vol. 26, N 6,— P. 730—733.
12. Prakash B. S., Meyer H. H. D., Schallenberger E., van de Wiel D. F. M. // J. Steroid Biochem.— 1987.— Vol. 28, N 6.— P. 623—627.
13. Rodbard D. // Analyt. Biochem.— 1978.— Vol. 90.— P. 1 — 12.
14. Sauer M. J., Foulkes J. A., Cookson A. D. // Steroids.— 1981,—Vol. 38, N 1,—P. 45—53.
Review
For citations:
Kolosova A.Yu., Yeremin S.A., Gavrilova Ye.M., Yegorov A.M. Polarization fluoroimmunoassays of progesterone. Problems of Endocrinology. 1994;40(4):48-51. (In Russ.) https://doi.org/10.14341/probl12147
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