An Electrochemical Detection of Metallothioneins at the Zeptomole Level in Nanolitre Volumes

Overview

Journal Sensors (Basel)

Publisher MDPI

Specialty Biotechnology

Date 2016 Nov 24

PMID 27879822

Citations 13

Authors

Vojtech Adam

Jiri Baloun

Ivo Fabrik

Libuse Trnkova

Rene Kizek

Affiliations

Soon will be listed here.

Abstract

An Electrochemical Detection of Metallothioneins at the Zeptomole Level in Nanolitre VolumesWe report on improvement of the adsorptive transfer stripping technique (AdTS) coupled with the differential pulse voltammetry Brdicka reaction to determine a thiol-protein. The current technique has been unable to generate reproducible results when analyzing very low sample volumes (nanolitres). This obstacle can be overcome technically by modifying the current transfer technique including cooling step of the adsorbed analyte. We tested the technique on determination of a promising tumour disease marker protein called metallothionein (MT). The detection limit (3 S/N) of MT was evaluated as 500 zeptomoles per 500 nL (1 pM) and the quantification limit (10 S/N) as 1,500 zeptomoles per 500 nL (3 pM). Further, the improved AdTS technique was utilized to analyze blood serum samples from patients with breast cancer. Based on the results obtained it can be concluded that the improved technique can be used to detect a thiolprotein in very low sample volumes and can also prevent interferences during the washing and transferring step.

Citing Articles

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A Determination of Metallothionein in Larvae of Freshwater Midges (Chironomus riparius) Using Brdicka Reaction.

Fabrik I, Ruferova Z, Hilscherova K, Adam V, Trnkova L, Kizek R Sensors (Basel). 2016; 8(7):4081-4094.

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Metallothionein modulation in relation to cadmium bioaccumulation and age-dependent sensitivity of Chironomus riparius larvae.

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References

Szpunar J . Bio-inorganic speciation analysis by hyphenated techniques. Analyst. 2000; 125(5):963-88. DOI: 10.1039/a909137h. View

Ferrarello C, Fernandez de la Campa M, Carrasco J, Sanz-Medel A . Speciation of metallothionein-like proteins of the mussel Mytilus edulis at basal levels by chromatographic separations coupled to quadrupole and double-focusing magnetic sector ICPMS. Anal Chem. 2001; 72(24):5874-80. DOI: 10.1021/ac0004411. View

Pale ek E, Fojta M . Detecting DNA hybridization and damage. Anal Chem. 2001; 73(3):74A-83A. DOI: 10.1021/ac0123936. View

Kizek R, Trnkova T, Palecek E . Determination of metallothionein at the femtomole level by constant current stripping chronopotentiometry. Anal Chem. 2001; 73(20):4801-7. DOI: 10.1021/ac010126u. View

Wang Z, Prange A . Use of surface-modified capillaries in the separation and characterization of metallothionein isoforms by capillary electrophoresis inductively coupled plasma mass spectrometry. Anal Chem. 2002; 74(3):626-31. DOI: 10.1021/ac010862y. View

Wolf C, Schaumloffel D, Richarz A, Prange A, Bratter P . CZE-ICP-MS separation of metallothioneins in human brain cytosols: comparability of electropherograms obtained from different sample matrices. Analyst. 2003; 128(6):576-80. DOI: 10.1039/b300030n. View

Masarik M, Kizek R, Kramer K, Billova S, Brazdova M, Vacek J . Application of avidin-biotin technology and adsorptive transfer stripping square-wave voltammetry for detection of DNA hybridization and avidin in transgenic avidin maize. Anal Chem. 2003; 75(11):2663-9. DOI: 10.1021/ac020788z. View

Luo D, Lan J, Zhou C, Luo C . Polarographic behavior of Co(II)-BSA or -HSA complex in the presence of a guanidine modifier. Anal Chem. 2003; 75(22):6346-50. DOI: 10.1021/ac0300643. View

Palecek E . Surface-attached molecular beacons light the way for DNA sequencing. Trends Biotechnol. 2004; 22(2):55-8. DOI: 10.1016/j.tibtech.2003.11.009. View

10.

Jelen F, Kourilova A, Pecinka P, Palecek E . Microanalysis of DNA by stripping transfer voltammetry. Bioelectrochemistry. 2004; 63(1-2):249-52. DOI: 10.1016/j.bioelechem.2003.10.021. View

11.

Palecek E, Masarik M, Kizek R, Kuhlmeier D, Hassmann J, Schulein J . Sensitive electrochemical determination of unlabeled MutS protein and detection of point mutations in DNA. Anal Chem. 2004; 76(19):5930-6. DOI: 10.1021/ac049474x. View

12.

Fojta M, Havran L, Kizek R, Billova S, Palecek E . Multiply osmium-labeled reporter probes for electrochemical DNA hybridization assays: detection of trinucleotide repeats. Biosens Bioelectron. 2004; 20(5):985-94. DOI: 10.1016/j.bios.2004.06.015. View

13.

Kizek R, Masarik M, Kramer K, Potesil D, Bailey M, Howard J . An analysis of avidin, biotin and their interaction at attomole levels by voltammetric and chromatographic techniques. Anal Bioanal Chem. 2005; 381(6):1167-78. DOI: 10.1007/s00216-004-3027-x. View

14.

Szpunar J . Advances in analytical methodology for bioinorganic speciation analysis: metallomics, metalloproteomics and heteroatom-tagged proteomics and metabolomics. Analyst. 2005; 130(4):442-65. DOI: 10.1039/b418265k. View

15.

Harrington C, Vidler D, Watts M, Hall J . Potential for using isotopically altered metalloproteins in species-specific isotope dilution analysis of proteins by HPLC coupled to inductively coupled plasma mass spectrometry. Anal Chem. 2005; 77(13):4034-41. DOI: 10.1021/ac050256t. View

16.

Legendre L, Bienvenue J, Roper M, Ferrance J, Landers J . A simple, valveless microfluidic sample preparation device for extraction and amplification of DNA from nanoliter-volume samples. Anal Chem. 2006; 78(5):1444-51. DOI: 10.1021/ac0516988. View

17.

Potesil D, Mikelova R, Adam V, Kizek R, Prusa R . Change of the protein p53 electrochemical signal according to its structural form - quick and sensitive distinguishing of native, denatured, and aggregated form of the "guardian of the genome". Protein J. 2006; 25(1):23-32. DOI: 10.1007/s10930-006-0014-4. View

18.

Du W, Fang Q, Fang Z . Microfluidic sequential injection analysis in a short capillary. Anal Chem. 2006; 78(18):6404-10. DOI: 10.1021/ac060714d. View

19.

Lorenz R, Edgar J, Jeffries G, Chiu D . Microfluidic and optical systems for the on-demand generation and manipulation of single femtoliter-volume aqueous droplets. Anal Chem. 2006; 78(18):6433-9. DOI: 10.1021/ac060748l. View

20.

Bertolini F, Shaked Y, Mancuso P, Kerbel R . The multifaceted circulating endothelial cell in cancer: towards marker and target identification. Nat Rev Cancer. 2006; 6(11):835-45. DOI: 10.1038/nrc1971. View