A rapid elisa method for the detection of sarcosine using pseudoperoxidase activity of gold nanoparticles

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60461373%3A22330%2F18%3A43916160" target="_blank" >RIV/60461373:22330/18:43916160 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216208:11310/18:10392588

Výsledek na webu

<a href="https://www.nanocon.eu/files/uploads/01/NANOCON2017_Proceedings_content.pdf" target="_blank" >https://www.nanocon.eu/files/uploads/01/NANOCON2017_Proceedings_content.pdf</a>

DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

A rapid elisa method for the detection of sarcosine using pseudoperoxidase activity of gold nanoparticles

Popis výsledku v původním jazyce

Sarcosine is an amino acid that is formed by methylation of glycine and is present in trace amounts in the body. Increased sarcosine concentrations in blood plasma and urine are associated with sarcosinemia and some other diseases such as prostate cancer. In addition, sarcosine may be administered as a drug in some psychiatric disorders. Primarily, sarcosine in biological fluids is analyzed using liquid chromatography techniques, but this technique is not suitable for routine analysis. For this purpose, sarcosine detection using the nanomedicine approach was proposed. In the presented test, Au20 gold nanoparticles (size 20-30 nm, zeta potential -38 mV, absorption maximum at 529 nm), which exhibit pseudoperoxidase activity and convert a suitable substrate such as TMB, ABTS (1 mM) to a distinctive color product, were used. In the following experiments, the Au20 nanoparticle was linked to the chicken anti-sarcosine antibody. Subsequently, the ELISA method for the determination of sarcosine was prepared. Firstly, the construct for direct detection of sarcosine bound to the antibody was prepared by both the direct and the indirect method (dependencies were strictly linear, R-2 = 0.996). The subsequent construct uses rabbit antibodies against the anti-sarcosine chicken antibody labelled with Au20 nanoparticle. The bound product is then monitored by the conversion of the TMB (655 nm) or ABTS (425 nm) substrate (the dependencies are strictly linear, R-2 = 0.998). To further increase the sensitivity of the method (for more than 90% of the original signal), an aggregation agent was added to the used Au20 nanoparticles, which further increased adherence of gold nanoparticles to the antibodies. This procedure succeeded in achieving subnanomolar detection limits for the amino acid sarcosine.

Název v anglickém jazyce

A rapid elisa method for the detection of sarcosine using pseudoperoxidase activity of gold nanoparticles

Popis výsledku anglicky

Sarcosine is an amino acid that is formed by methylation of glycine and is present in trace amounts in the body. Increased sarcosine concentrations in blood plasma and urine are associated with sarcosinemia and some other diseases such as prostate cancer. In addition, sarcosine may be administered as a drug in some psychiatric disorders. Primarily, sarcosine in biological fluids is analyzed using liquid chromatography techniques, but this technique is not suitable for routine analysis. For this purpose, sarcosine detection using the nanomedicine approach was proposed. In the presented test, Au20 gold nanoparticles (size 20-30 nm, zeta potential -38 mV, absorption maximum at 529 nm), which exhibit pseudoperoxidase activity and convert a suitable substrate such as TMB, ABTS (1 mM) to a distinctive color product, were used. In the following experiments, the Au20 nanoparticle was linked to the chicken anti-sarcosine antibody. Subsequently, the ELISA method for the determination of sarcosine was prepared. Firstly, the construct for direct detection of sarcosine bound to the antibody was prepared by both the direct and the indirect method (dependencies were strictly linear, R-2 = 0.996). The subsequent construct uses rabbit antibodies against the anti-sarcosine chicken antibody labelled with Au20 nanoparticle. The bound product is then monitored by the conversion of the TMB (655 nm) or ABTS (425 nm) substrate (the dependencies are strictly linear, R-2 = 0.998). To further increase the sensitivity of the method (for more than 90% of the original signal), an aggregation agent was added to the used Au20 nanoparticles, which further increased adherence of gold nanoparticles to the antibodies. This procedure succeeded in achieving subnanomolar detection limits for the amino acid sarcosine.

Druh

D - Stať ve sborníku

CEP obor

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OECD FORD obor

21002 - Nano-processes (applications on nano-scale); (biomaterials to be 2.9)

Projekt

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Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2018

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název statě ve sborníku

Nanocon 2017

ISBN

978-80-87294-81-9

ISSN

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e-ISSN

neuvedeno

Počet stran výsledku

6

Strana od-do

524-530

Název nakladatele

Tanger s.r.o.

Místo vydání

Ostrava

Místo konání akce

Brno

Datum konání akce

18. 10. 2017

Typ akce podle státní příslušnosti

WRD - Celosvětová akce

Kód UT WoS článku

000452823300086