Michaela Vitkova
EnzymeChemistryCaseinFormononetinGenistinGenisteinIsoflavonoidBiochanin AIsoflavonesImmunoassayRutaOnoninDaidzeinRutaceaeHaptenChromatographyBiochemistryBiologyBovine serum albuminPolyclonal antibodies
7Publications
5H-index
58Citations
Publications 7
Newest
#1Michaela VitkovaH-Index: 5
#2Z. MackovaH-Index: 2
Last. Oldřich LapčíkH-Index: 14
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The development of polyclonal antibody-based enzyme-linked immunosorbent assays (ELISA) for the determination of individual isoflavones, i.e. daidzein, genistein and biochanin and their homologues, is presented in this work. Isoflavone conjugates with bovine serum albumin were used as immunogens, coupled at the position C 7 and C 4' via a carboxy methyl spacer. The developed ELISAs are highly specific, I 5 0 values of the standard curves range between 0.3-1.2 ng/ml. The cross reactivities to oth...
2 CitationsSource
#1Michaela VitkovaH-Index: 5
#2Pavel RauchH-Index: 15
Last. Ladislav FukalH-Index: 9
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Polyclonal antibodies were raised against six immunogens (three native and three thermally treated casein fractions: α+β-casein, K-casein and whole casein). Using these antibodies the procedures of an indirect competitiveenzyme immunoassay (ELISA) were constructed, optimised and characterised for determination of each immunogen. It was found that ELISA of caseins is very specific without any interferences of whey proteins and with proportionally low cross-interactions between caseins. Detection ...
2 CitationsSource
#1Radka KoblovskáH-Index: 5
#2Zuzana MackovaH-Index: 4
Last. Oldřich LapčíkH-Index: 14
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Enzyme-linked immunosorbent assays in combination with semi-preparative high-performance liquid chromatography (HPLC) and analytical HPLC with mass spectroscopy in the selective ion monitoring mode were used for the determination of selected isoflavones, daidzein, genistein, biochanin A and their homologues, in 20 representatives of the Rutaceae family. Species belonging to five genera were studied, namely Citrus, Fortunella, Poncirus, Ruta and Severinia. The enzyme immunoassays used were based ...
12 CitationsSource
#1O. LapčíkH-Index: 1
#1Oldřich LapčíkH-Index: 14
Last. Bořivoj Klejdus (H.I., S.I.: University of Agriculture, Faisalabad)H-Index: 36
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Extracts from Arabidopsis thaliana leaves and inflorescence stalks and from Lepidium sativa (Brassicaceae) seedlings were analysed by HPLC-MS-SIM and by five isoflavonoid-specific ELISA methods after the HPLC fractionation of samples, in order to determine presence of isoflavonoids. Individual ELISAs were specific for daidzein, genistein, biochanin A and for their derivatives substituted either at the 4′- or at the 7- positions. Both analytical approaches revealed homologous spectra of isoflavon...
18 CitationsSource
#1Oldřich LapčíkH-Index: 14
#2Michaela VitkovaH-Index: 5
Last. Herman Adlercreutz (UH: University of Helsinki)H-Index: 99
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Abstract Two variants of immunoassay for the determination of biochanin A (5,7-dihydroxy 4′-methoxy isoflavone), i.e., a radioimmunoassay (RIA) and an indirect ELISA, have been developed and evaluated. Both methods employ the same rabbit antiserum to a 7- O -carboxymethyl-5-hydroxy-4′-methoxyisoflavone-bovine serum albumin (BSA) conjugate. A 125 I-labeled hapten-tyrosine methyl ester (TME) conjugate was used as a radioligand for the RIA. The indirect ELISA uses immunogen-coated microtitration pl...
13 CitationsSource
#1Michaela VitkovaH-Index: 5
#2Zuzana MackovaH-Index: 4
Last. Oldrich LapcikH-Index: 5
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Competitive enzyme-linked immunosorbent assays (ELISA) for the determination of isoflavones daidzein, genistein and biochanin were developed. Isoflavone conjugates coupled at positions C7 and C4' via a carboxymethyl spacer to bovine serum albumin were used as immunogens and also as coating antigens. Assay conditions, including concentrations of antisera and coating antigens, were optimized and the effect of reaction time, various blocking agents and organic solvents was investigated. The develop...
6 Citations
#1Oldřich LapčíkH-Index: 14
#2Jan StursaH-Index: 21
Last. Jitka MoravcováH-Index: 8
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Abstract 3- O -Carboxymethylcoumestrol was prepared as the hapten for immunoassay by a partial alkylation of coumestrol with ethyl chloroacetate in acetone alkalized with potassium carbonate. 3- O -Ethoxycarbonylmethylcoumestrol was separated by column chromatography and finally was hydrolyzed with formic acid. 1 H and 13 C NMR data (APT, COSY, HMQC, and HMBC) revealed that the reaction was regioselective, as 3- O -ethoxycarboxymethylcoumestrol was the only monosubstituted derivative. The hapten...
8 CitationsSource
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