Development of amperometric immunosensor for diagnosis of
albuminuria based on screen-printed electrodes

D 2018

Development of amperometric immunosensor for diagnosis of albuminuria based on screen-printed electrodes

MIKUŠOVÁ, Zuzana, Matěj PASTUCHA, Zdeněk FARKA, Veronika POLÁCHOVÁ, Petr SKLÁDAL et. al.

Základní údaje

Originální název

Development of amperometric immunosensor for diagnosis of albuminuria based on screen-printed electrodes

Autoři

MIKUŠOVÁ, Zuzana, Matěj PASTUCHA, Zdeněk FARKA, Veronika POLÁCHOVÁ a Petr SKLÁDAL

Vydání

Brno, XVIII. Workshop of Biophysical Chemists and Electrochemists, od s. 33-34, 2 s. 2018

Nakladatel

Masarykova univerzita

Další údaje

Jazyk

angličtina

Typ výsledku

Stať ve sborníku

Obor

10406 Analytical chemistry

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Forma vydání

elektronická verze "online"

Odkazy

URL

Organizační jednotka

Středoevropský technologický institut

ISBN

978-80-210-9029-3

Klíčová slova anglicky

Biosensor; Amperometry; Sandwich assay; Human serum albumin; Albuminuria

Změněno: 15. 9. 2018 20:05, doc. Mgr. Zdeněk Farka, Ph.D.

Anotace

V originále

Human serum albumin (HSA) is a plasma protein which plays an important role in transporting ligands to specific targets. Increased presence of HSA in urine (microalbuminuria) is considered as an indication of diabetic nephropathy or glomerulonephritis. The amount of HSA in 24-h urine specimens typical for microalbuminuria ranges from 30 to 300 mg. Nowadays, there are various immunoassays for detection of HSA such as enzyme linked immunosorbent assay (ELISA). These methods provide high sensitivity; however, they are time consuming, complicated and require expensive instrumentation. An appropriate solution to overcome these disadvantages is the use of electrochemical biosensors that offer operational simplicity, high sensitivity, short analysis times and miniaturization possibilities for point-of-care diagnosis. The electrochemical biosensors are typically based on screen-printed electrodes (SPEs) due to their low cost, portability and mass production capabilities. We have developed an amperometric immunosensor for rapid detection of HSA in urine. Anti-HSA antibody was immobilized to the surface of electrodes that allowed specific capture of HSA. The detection was based on sandwich immunoassay principle where the secondary antibody conjugated with horseradish peroxidase (HRP) was binding to surface captured immunocomplex Ab1-HSA. HRP as label provides oxidation of 3,3´,5,5´-tetramethylbenzidine (TMB) in presence of H2O2. Alternatively, we used Prussian blue catalytic nanoparticles as convenient enzyme replacement. Non-specific binding was evaluated by incubating the sensor with BSA instead of HSA. We compered SPEs based on gold and carbon (including carbon nanotubes) working electrodes. So far, the lowest concentration of detected HSA was 10 μg·mL−1. The actual limit of detection is useful for practical analysis and point-of-care diagnosis. This method has proved to be suitable for diagnostics of microalbuminuria. Optimization focused on better detection limit and to reduce, respectively eliminate non-specific binding is in progress.

Návaznosti

LQ1601, projekt VaV

Název: CEITEC 2020 (Akronym: CEITEC2020)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, CEITEC 2020

Citovat

MIKUŠOVÁ, Zuzana, Matěj PASTUCHA, Zdeněk FARKA, Veronika POLÁCHOVÁ a Petr SKLÁDAL. Development of amperometric immunosensor for diagnosis of albuminuria based on screen-printed electrodes. Online. In Libuše Trnková. XVIII. Workshop of Biophysical Chemists and Electrochemists. Brno: Masarykova univerzita, 2018, s. 33-34. ISBN 978-80-210-9029-3.

@inproceedings{1436497,
   author = {Mikušová, Zuzana and Pastucha, Matěj and Farka, Zdeněk and Poláchová, Veronika and Skládal, Petr},
   address = {Brno},
   booktitle = {XVIII. Workshop of Biophysical Chemists and Electrochemists},
   editor = {Libuše Trnková},
   keywords = {Biosensor; Amperometry; Sandwich assay; Human serum albumin; Albuminuria},
   howpublished = {elektronická verze "online"},
   language = {eng},
   location = {Brno},
   isbn = {978-80-210-9029-3},
   pages = {33-34},
   publisher = {Masarykova univerzita},
   title = {Development of amperometric immunosensor for diagnosis of albuminuria based on screen-printed electrodes},
   url = {http://www.sci.muni.cz/~labifel/files/soubory/sbornik_2018.pdf#page=33},
   year = {2018}
}

TY  - JOUR
ID  - 1436497
AU  - Mikušová, Zuzana - Pastucha, Matěj - Farka, Zdeněk - Poláchová, Veronika - Skládal, Petr
PY  - 2018
TI  - Development of amperometric immunosensor for diagnosis of albuminuria based on screen-printed electrodes
PB  - Masarykova univerzita
CY  - Brno
SN  - 9788021090293
KW  - Biosensor
KW  - Amperometry
KW  - Sandwich assay
KW  - Human serum albumin
KW  - Albuminuria
UR  - http://www.sci.muni.cz/~labifel/files/soubory/sbornik_2018.pdf#page=33
N2  - Human serum albumin (HSA) is a plasma protein which plays an important role in transporting ligands to specific targets. Increased presence of HSA in urine (microalbuminuria) is considered as an indication of diabetic nephropathy or glomerulonephritis. The amount of HSA in 24-h urine specimens typical for microalbuminuria ranges from 30 to 300 mg. Nowadays, there are various immunoassays for detection of HSA such as enzyme linked immunosorbent assay (ELISA). These methods provide high sensitivity; however, they are time consuming, complicated and require expensive instrumentation. An appropriate solution to overcome these disadvantages is the use of electrochemical biosensors that offer operational simplicity, high sensitivity, short analysis times and miniaturization possibilities for point-of-care diagnosis. The electrochemical biosensors are typically based on screen-printed electrodes (SPEs) due to their low cost, portability and mass production capabilities. We have developed an amperometric immunosensor for rapid detection of HSA in urine. Anti-HSA antibody was immobilized to the surface of electrodes that allowed specific capture of HSA. The detection was based on sandwich immunoassay principle where the secondary antibody conjugated with horseradish peroxidase (HRP) was binding to surface captured immunocomplex Ab1-HSA. HRP as label provides oxidation of 3,3´,5,5´-tetramethylbenzidine (TMB) in presence of H2O2. Alternatively, we used Prussian blue catalytic nanoparticles as convenient enzyme replacement. Non-specific binding was evaluated by incubating the sensor with BSA instead of HSA. We compered SPEs based on gold and carbon (including carbon nanotubes) working electrodes. So far, the lowest concentration of detected HSA was 10 μg·mL−1. The actual limit of detection is useful for practical analysis and point-of-care diagnosis. This method has proved to be suitable for diagnostics of microalbuminuria. Optimization focused on better detection limit and to reduce, respectively eliminate non-specific binding is in progress.
ER  -

MIKUŠOVÁ, Zuzana, Matěj PASTUCHA, Zdeněk FARKA, Veronika POLÁCHOVÁ a Petr SKLÁDAL. Development of amperometric immunosensor for diagnosis of albuminuria based on screen-printed electrodes. Online. In Libuše Trnková. \textit{XVIII. Workshop of Biophysical Chemists and Electrochemists}. Brno: Masarykova univerzita, 2018, s.~33-34. ISBN~978-80-210-9029-3.

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