Immunoextraction of zinc proteins from human plasma using
chicken yolk antibodies immobilized onto paramagnetic particles
and their electrophoretic analysis

J 2012

Immunoextraction of zinc proteins from human plasma using chicken yolk antibodies immobilized onto paramagnetic particles and their electrophoretic analysis

KŘÍŽKOVÁ, Soňa, Markéta RYVOLOVÁ, David HYNEK, Tomáš ECKSCHLAGER, Petr HODEK et. al.

Základní údaje

Originální název

Immunoextraction of zinc proteins from human plasma using chicken yolk antibodies immobilized onto paramagnetic particles and their electrophoretic analysis

Autoři

KŘÍŽKOVÁ, Soňa (203 Česká republika, garant), Markéta RYVOLOVÁ (203 Česká republika), David HYNEK (203 Česká republika), Tomáš ECKSCHLAGER (203 Česká republika), Petr HODEK (203 Česká republika), Michal MASAŘÍK (203 Česká republika, domácí), Vojtěch ADAM (203 Česká republika) a René KIZEK (203 Česká republika)

Vydání

Electrophoresis, 2012, 0173-0835

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10405 Electrochemistry

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 3.261

Kód RIV

RIV/00216224:14110/12:00060406

Organizační jednotka

Lékařská fakulta

DOI

http://dx.doi.org/10.1002/elps.201100638

UT WoS

000305792500018

Klíčová slova anglicky

Electrochemistry; Magnetic particles; Metallomics; Zincome

Příznaky

Mezinárodní význam

Změněno: 16. 4. 2013 15:47, Ing. Mgr. Věra Pospíšilíková

Anotace

V originále

Zinc(II) as the only transition metal lacking redox activity is an essential part of approximately 10% proteins as a cofactor of these proteins. Considering the fact that there are numerous zinc(II) containing proteins, proteomics and metallomics studies aimed on them require accurate methods for preparation of real biological samples prior to their subsequent analysis using 2DE and MS. For this purpose, we suggested a new method based on chicken anti-zinc antibodies and magnetizable particles. Antibodies were covalently immobilized to the surface of paramagnetic beads activated with tosyl group. Binding of the antibody to the beads was confirmed by secondary anti-chicken antibody conjugated with horseradish peroxidase. The immunoextraction conditions, such as concentration of the beads, time of immunoextraction, pH and composition of the elution buffer, and time of extraction were optimized. Subsequently, zinc proteins were extracted from human plasma and total concentration of zinc was monitored by electrochemical detection in the extracts. Under optimal conditions it was possible to monitor the proteins and zinc removal from the sample by chip CE, SDS-PAGE, and indirectly using electrochemistry.

Citovat

KŘÍŽKOVÁ, Soňa, Markéta RYVOLOVÁ, David HYNEK, Tomáš ECKSCHLAGER, Petr HODEK, Michal MASAŘÍK, Vojtěch ADAM a René KIZEK. Immunoextraction of zinc proteins from human plasma using chicken yolk antibodies immobilized onto paramagnetic particles and their electrophoretic analysis. Electrophoresis. 2012, roč. 33, č. 12, s. 1824-1832. ISSN 0173-0835. Dostupné z: https://dx.doi.org/10.1002/elps.201100638.

@article{987331,
   author = {Křížková, Soňa and Ryvolová, Markéta and Hynek, David and Eckschlager, Tomáš and Hodek, Petr and Masařík, Michal and Adam, Vojtěch and Kizek, René},
   article_number = {12},
   doi = {http://dx.doi.org/10.1002/elps.201100638},
   keywords = {Electrochemistry; Magnetic particles; Metallomics; Zincome},
   language = {eng},
   issn = {0173-0835},
   journal = {Electrophoresis},
   title = {Immunoextraction of zinc proteins from human plasma using chicken yolk antibodies immobilized onto paramagnetic particles and their electrophoretic analysis},
   volume = {33},
   year = {2012}
}

TY  - JOUR
ID  - 987331
AU  - Křížková, Soňa - Ryvolová, Markéta - Hynek, David - Eckschlager, Tomáš - Hodek, Petr - Masařík, Michal - Adam, Vojtěch - Kizek, René
PY  - 2012
TI  - Immunoextraction of zinc proteins from human plasma using chicken yolk antibodies immobilized onto paramagnetic particles and their electrophoretic analysis
JF  - Electrophoresis
VL  - 33
IS  - 12
SP  - 1824-1832
EP  - 1824-1832
SN  - 01730835
KW  - Electrochemistry
KW  - Magnetic particles
KW  - Metallomics
KW  - Zincome
N2  - Zinc(II) as the only transition metal lacking redox activity is an essential part of approximately 10% proteins as a cofactor of these proteins. Considering the fact that there are numerous zinc(II) containing proteins, proteomics and metallomics studies aimed on them require accurate methods for preparation of real biological samples prior to their subsequent analysis using 2DE and MS. For this purpose, we suggested a new method based on chicken anti-zinc antibodies and magnetizable particles. Antibodies were covalently immobilized to the surface of paramagnetic beads activated with tosyl group. Binding of the antibody to the beads was confirmed by secondary anti-chicken antibody conjugated with horseradish peroxidase. The immunoextraction conditions, such as concentration of the beads, time of immunoextraction, pH and composition of the elution buffer, and time of extraction were optimized. Subsequently, zinc proteins were extracted from human plasma and total concentration of zinc was monitored by electrochemical detection in the extracts. Under optimal conditions it was possible to monitor the proteins and zinc removal from the sample by chip CE, SDS-PAGE, and indirectly using electrochemistry.
ER  -

KŘÍŽKOVÁ, Soňa, Markéta RYVOLOVÁ, David HYNEK, Tomáš ECKSCHLAGER, Petr HODEK, Michal MASAŘÍK, Vojtěch ADAM a René KIZEK. Immunoextraction of zinc proteins from human plasma using chicken yolk antibodies immobilized onto paramagnetic particles and their electrophoretic analysis. \textit{Electrophoresis}. 2012, roč.~33, č.~12, s.~1824-1832. ISSN~0173-0835. Dostupné z: https://dx.doi.org/10.1002/elps.201100638.

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