Publication detail

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

KŘÍŽKOVÁ, S. VACULOVIČOVÁ, M. HYNEK, D. ECKSCHLAGER, T. HODEK, P. MASAŘÍK, M. ADAM, V. KIZEK, R.

Original Title

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

Type

journal article in Web of Science

Language

English

Original Abstract

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 (6–18 g/mL of the sample), time of immunoextraction (6–34 min), pH and composition of the elution buffer, and time of extraction (48–300 s) 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.

Keywords

Electrochemistry; Magnetic particles; Metallomics; Zincome

Authors

KŘÍŽKOVÁ, S.; VACULOVIČOVÁ, M.; HYNEK, D.; ECKSCHLAGER, T.; HODEK, P.; MASAŘÍK, M.; ADAM, V.; KIZEK, R.

RIV year

2012

Released

5. 6. 2012

ISBN

0173-0835

Periodical

Electrophoresis

Year of study

33

Number

12

State

Federal Republic of Germany

Pages from

1824

Pages to

1832

Pages count

9

BibTex

@article{BUT92846,
  author="Soňa {Křížková} and Markéta {Vaculovičová} and David {Hynek} and Tomáš {Eckschlager} and Petr {Hodek} and Michal {Masařík} and Vojtěch {Adam} and René {Kizek}",
  title="Immunoextraction of zinc proteins from human plasma using chicken yolk antibodies immobilized onto paramagnetic particles and their electrophoretic analysis",
  journal="Electrophoresis",
  year="2012",
  volume="33",
  number="12",
  pages="1824--1832",
  doi="10.1002/elps.201100638",
  issn="0173-0835"
}