Isolation of metallothionein from cells derived from aggressive
form of high-grade prostate carcinoma using paramagnetic
antibody-modified microbeads off-line coupled with
electrochemical and electrophoretic analysis

J 2011

Isolation of metallothionein from cells derived from aggressive form of high-grade prostate carcinoma using paramagnetic antibody-modified microbeads off-line coupled with electrochemical and electrophoretic analysis

MASAŘÍK, Michal, Jaromír GUMULEC, Markéta SZTALMACHOVÁ, Marián HLAVNA, Petr BABELA et. al.

Základní údaje

Originální název

Isolation of metallothionein from cells derived from aggressive form of high-grade prostate carcinoma using paramagnetic antibody-modified microbeads off-line coupled with electrochemical and electrophoretic analysis

Autoři

MASAŘÍK, Michal (203 Česká republika, garant, domácí), Jaromír GUMULEC (203 Česká republika, domácí), Markéta SZTALMACHOVÁ (203 Česká republika, domácí), Marián HLAVNA (703 Slovensko, domácí), Petr BABELA (203 Česká republika), Soňa KŘÍŽKOVÁ (203 Česká republika), Markéta RYVOLOVÁ (203 Česká republika), Michal JURAJDA (203 Česká republika, domácí), Jiří SOCHOR (203 Česká republika), Vojtěch ADAM (203 Česká republika) a René KIZEK (203 Česká republika, domácí)

Vydání

Electrophoresis, 2011, 0173-0835

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10600 1.6 Biological sciences

Stát vydavatele

Německo

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 3.303

Kód RIV

RIV/00216224:14110/11:00050260

Organizační jednotka

Lékařská fakulta

DOI

http://dx.doi.org/10.1002/elps.201100301

UT WoS

000298587900014

Klíčová slova anglicky

Immunoprecipitation; Metallothionein; Microparticles; Microseparation; Prostate cancer

Příznaky

Mezinárodní význam

Změněno: 20. 12. 2011 14:19, Mgr. Michal Petr

Anotace

V originále

Prostate cancer with altered zinc(II) cell metabolism is the second most frequently diagnosed cancer in developed countries. The alterations of zinc(II) metabolism can influence metabolism of other metal ions and can also be associated with the expression and translation of metal-binding proteins including metallothioneins. The aim of this article was to optimize immunoseparation protocol based on paramagnetic beads conjugated with protein G for the isolation of metallothionein. Isolated metallothionein was determined by differential pulse voltammetry Brdicka reaction and SDS-PAGE. Optimal conditions: antigen-binding time - 60 min, temperature - 70 C, and buffer composition and pH - acetate buffer, pH 4,3, were determined. Under the optimized conditions, lysates from 22Rv1 prostate cancer cells treated with various concentrations of cadmium(II) and copper(II) ions were analyzed. We observed strong correlation in all experimental groups and all lysate types between metallothionein concentration related to viability and concentration of copper(II) ions and cadmium(II) ions in medium. Moreover, the results were compared with standard sample preparation as heat treatment and SDS-PAGE analysis.

Návaznosti

GP301/09/P436, projekt VaV

Název: Analýza metalothioneinu u karcinomu prostaty na úrovni DNA, RNA a proteinu.

Investor: Grantová agentura ČR, Analýza metalothioneinu u karcinomu prostaty na úrovni DNA, RNA a proteinu

NS10200, projekt VaV

Název: Nádorové markery, jejich stanovení a korelace s karcinomem prostaty

Investor: Ministerstvo zdravotnictví ČR, Nádorové markery, jejich stanovení a korelace s karcinomem prostaty

Citovat

MASAŘÍK, Michal, Jaromír GUMULEC, Markéta SZTALMACHOVÁ, Marián HLAVNA, Petr BABELA, Soňa KŘÍŽKOVÁ, Markéta RYVOLOVÁ, Michal JURAJDA, Jiří SOCHOR, Vojtěch ADAM a René KIZEK. Isolation of metallothionein from cells derived from aggressive form of high-grade prostate carcinoma using paramagnetic antibody-modified microbeads off-line coupled with electrochemical and electrophoretic analysis. Electrophoresis. 2011, roč. 32, č. 24, s. 3576–3588. ISSN 0173-0835. Dostupné z: https://dx.doi.org/10.1002/elps.201100301.

@article{963512,
   author = {Masařík, Michal and Gumulec, Jaromír and Sztalmachová, Markéta and Hlavna, Marián and Babela, Petr and Křížková, Soňa and Ryvolová, Markéta and Jurajda, Michal and Sochor, Jiří and Adam, Vojtěch and Kizek, René},
   article_number = {24},
   doi = {http://dx.doi.org/10.1002/elps.201100301},
   keywords = {Immunoprecipitation; Metallothionein; Microparticles; Microseparation; Prostate cancer},
   language = {eng},
   issn = {0173-0835},
   journal = {Electrophoresis},
   title = {Isolation of metallothionein from cells derived from aggressive form of high-grade prostate carcinoma using paramagnetic antibody-modified microbeads off-line coupled with electrochemical and electrophoretic analysis},
   volume = {32},
   year = {2011}
}

TY  - JOUR
ID  - 963512
AU  - Masařík, Michal - Gumulec, Jaromír - Sztalmachová, Markéta - Hlavna, Marián - Babela, Petr - Křížková, Soňa - Ryvolová, Markéta - Jurajda, Michal - Sochor, Jiří - Adam, Vojtěch - Kizek, René
PY  - 2011
TI  - Isolation of metallothionein from cells derived from aggressive form of high-grade prostate carcinoma using paramagnetic antibody-modified microbeads off-line coupled with electrochemical and electrophoretic analysis
JF  - Electrophoresis
VL  - 32
IS  - 24
SP  - 3576–3588
EP  - 3576–3588
SN  - 01730835
KW  - Immunoprecipitation
KW  - Metallothionein
KW  - Microparticles
KW  - Microseparation
KW  - Prostate cancer
N2  - Prostate cancer with altered zinc(II) cell metabolism is the second most frequently diagnosed cancer in developed countries. The alterations of zinc(II) metabolism can influence metabolism of other metal ions and can also be associated with the expression and translation of metal-binding proteins including metallothioneins. The aim of this article was to optimize immunoseparation protocol based on paramagnetic beads conjugated with protein G for the isolation of metallothionein. Isolated metallothionein was determined by differential pulse voltammetry Brdicka reaction and SDS-PAGE. Optimal conditions: antigen-binding time - 60 min, temperature - 70 C, and buffer composition and pH - acetate buffer, pH 4,3, were determined. Under the optimized conditions, lysates from 22Rv1 prostate cancer cells treated with various concentrations of cadmium(II) and copper(II) ions were analyzed. We observed strong correlation in all experimental groups and all lysate types between metallothionein concentration related to viability and concentration of copper(II) ions and cadmium(II) ions in medium. Moreover, the results were compared with standard sample preparation as heat treatment and SDS-PAGE analysis.
ER  -

MASAŘÍK, Michal, Jaromír GUMULEC, Markéta SZTALMACHOVÁ, Marián HLAVNA, Petr BABELA, Soňa KŘÍŽKOVÁ, Markéta RYVOLOVÁ, Michal JURAJDA, Jiří SOCHOR, Vojtěch ADAM a René KIZEK. Isolation of metallothionein from cells derived from aggressive form of high-grade prostate carcinoma using paramagnetic antibody-modified microbeads off-line coupled with electrochemical and electrophoretic analysis. \textit{Electrophoresis}. 2011, roč.~32, č.~24, s.~3576–3588. ISSN~0173-0835. Dostupné z: https://dx.doi.org/10.1002/elps.201100301.

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