Detail publikace

Paramagnetic antibody-modified microparticles coupled with voltammetry as a tool for isolation and detection of metallothionen as a bioindicator of metal pollution

TRNKOVÁ, L. HÚSKA, D. ŠKUTKOVÁ, H. BEKLOVÁ, M. HUBÁLEK, J. ADAM, V. PROVAZNÍK, I. KIZEK, R.

Originální název

Paramagnetic antibody-modified microparticles coupled with voltammetry as a tool for isolation and detection of metallothionen as a bioindicator of metal pollution

Český název

Paramagnetic antibody-modified microparticles coupled with voltammetry as a tool for isolation and detection of metallothionen as a bioindicator of metal pollution

Anglický název

Paramagnetic antibody-modified microparticles coupled with voltammetry as a tool for isolation and detection of metallothionen as a bioindicator of metal pollution

Typ

článek v časopise

Jazyk

en

Originální abstrakt

Low-molecular mass proteins rich in cysteines called metallothioneins (MT) can be considered as markers for the pollution of the environment by metals. Here, we report on suggestion for an automated procedure for the isolation of MT followed by voltammetric analysis. Primarily, we optimized the automated detection of MTusing an electrochemical analyser. It was found that the most sensitive and repeatable analyses are obtained at a temperature of 4C for the supporting electrolyte. Further, we optimized experimental conditions for the isolation of MT by using antibody-linked paramagnetic microparticles. Under the optimal conditions (4 h long interaction between the microparticles and MT), the microparticles were tested on isolation of various amounts of MT. The lowest isolated amount of MT by antibody-linked paramagnetic microparticles was 5 mg ml of MT (50 ng). The automated procedure of MT isolation was further tested on isolation of MT from guppy fish (Poecilia reticulata) treated with silver(I) ions (50 mM AgNO3). The whole process lasted less than five hours and was fully automated. We attempted to correlate these results with the standard method for MT isolation. The correlation coefficient is 0.9901, which confirms that results are in good agreement. Moreover, the concentration of silver ions in tissues of fish treated with Ag(I) ions was determined by high performance liquid chromatography with electrochemical detection.

Český abstrakt

Low-molekulová hmotnost proteinů bohatých na cysteiny tzv. metallothioneins (MT) lze považovat za ukazatele znečištění životního prostředí kovy. Zde zpráva o návrh na automatizovaný postup pro izolaci MT následně voltametrické analýzy. V prvé řadě jsme optimalizovali automatickou detekci MTusing analyzátoru electrochemical. Bylo zjištěno, že nejcitlivější a opakovatelné analýzy jsou získány při teplotě 4C pro základní elektrolyt. Dále jsme optimalizované experimentálních podmínek pro izolaci MT pomocí protilátky vázané na paramagnetické mikročástice. Za optimálních podmínek (4 h dlouho interakce mezi mikročástice a MT), byly testovány na mikročástice izolaci různých množství MT. Nejnižší izolované množství MT v protilátek vázaných paramagnetické mikročástice je 5 mg ml MT (50 ng).Automatického postupu izolace MT byl dále testován na izolaci MT z ryb Guppy (Poecilia reticulata) léčených stříbrný (I) ionty (50 mM AgNO3). Celý proces trval méně než pět hodin a byl plně automatizován. Pokusili jsme se tyto výsledky korelovat se standardní metoda pro izolaci MT.Korelační koeficient je 0,9901, což potvrzuje, že výsledky jsou v dobré shodě. Kromě toho byla koncentrace stříbrných iontů ve tkáních ryb léčeny ionty Ag (I) stanoví vysokoúčinnou kapalinovou chromatografií s detekcí elektrochemickou.

Anglický abstrakt

Low-molecular mass proteins rich in cysteines called metallothioneins (MT) can be considered as markers for the pollution of the environment by metals. Here, we report on suggestion for an automated procedure for the isolation of MT followed by voltammetric analysis. Primarily, we optimized the automated detection of MTusing an electrochemical analyser. It was found that the most sensitive and repeatable analyses are obtained at a temperature of 4C for the supporting electrolyte. Further, we optimized experimental conditions for the isolation of MT by using antibody-linked paramagnetic microparticles. Under the optimal conditions (4 h long interaction between the microparticles and MT), the microparticles were tested on isolation of various amounts of MT. The lowest isolated amount of MT by antibody-linked paramagnetic microparticles was 5 mg ml of MT (50 ng). The automated procedure of MT isolation was further tested on isolation of MT from guppy fish (Poecilia reticulata) treated with silver(I) ions (50 mM AgNO3). The whole process lasted less than five hours and was fully automated. We attempted to correlate these results with the standard method for MT isolation. The correlation coefficient is 0.9901, which confirms that results are in good agreement. Moreover, the concentration of silver ions in tissues of fish treated with Ag(I) ions was determined by high performance liquid chromatography with electrochemical detection.

Klíčová slova

protilátka, mikročástice, voltammetrie, metallothionen, znečištění

Rok RIV

2011

Vydáno

05.10.2011

Strany od

2763

Strany do

2769

Strany počet

7

BibTex


@article{BUT73809,
  author="Libuše {Trnková} and Dalibor {Húska} and Helena {Škutková} and Miroslava {Beklová} and Jaromír {Hubálek} and Vojtěch {Adam} and Ivo {Provazník} and René {Kizek}",
  title="Paramagnetic antibody-modified microparticles coupled with voltammetry as a tool for isolation and detection of metallothionen as a bioindicator of metal pollution",
  annote="Low-molecular mass proteins rich in cysteines called metallothioneins (MT) can be considered as markers for the pollution of the environment by metals. Here, we report on suggestion for an automated procedure for the isolation of MT followed by voltammetric analysis. Primarily, we optimized the automated detection of MTusing an electrochemical analyser. It was found that the most sensitive and repeatable analyses are obtained at a temperature of 4C for the supporting electrolyte. Further, we optimized experimental conditions for the isolation of MT by using antibody-linked paramagnetic microparticles. Under the optimal conditions (4 h long interaction between the microparticles and MT), the microparticles were tested on isolation of various amounts of MT. The lowest isolated amount of MT by antibody-linked paramagnetic microparticles was 5 mg ml of MT
(50 ng). The automated procedure of MT isolation was further tested on isolation of MT from guppy fish (Poecilia reticulata) treated with silver(I) ions (50 mM AgNO3). The whole process lasted less than five hours and was fully automated. We attempted to correlate these results with the standard method for MT isolation. The correlation coefficient is 0.9901, which confirms that results are in good agreement. Moreover, the concentration of silver ions in tissues of fish treated with Ag(I) ions was determined by high performance liquid chromatography with electrochemical detection.",
  chapter="73809",
  number="13",
  volume="2011",
  year="2011",
  month="october",
  pages="2763--2769",
  type="journal article"
}