Novel Fitc-Labeled Igy Antibody: Fluorescence Imaging Toxoplasma Gondii In Vitro

Abstract Toxoplasmosis is caused by T. gondii and can create serious health problems in humans and also worldwide economic harm. Because of the clinical and veterinary importance of toxoplasmosis, its timely and accurate diagnosis has a major impact on disease-fighting strategies. T. gondii surface...

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Main Authors: Mehtap Sert, Rabia Cakir Koc, Yasemin Budama Kilinc
Format: Article
Language:English
Published: Nature Publishing Group 2017-04-01
Series:Scientific Reports
Online Access:https://doi.org/10.1038/s41598-017-00930-1
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spelling doaj-31414e568a7448d18e5e37fbc9df38662020-12-08T02:05:18ZengNature Publishing GroupScientific Reports2045-23222017-04-01711710.1038/s41598-017-00930-1Novel Fitc-Labeled Igy Antibody: Fluorescence Imaging Toxoplasma Gondii In VitroMehtap Sert0Rabia Cakir Koc1Yasemin Budama Kilinc2Department of Bioengineering, Faculty of Chemical and Metallurgical Engineering, Yildiz Technical UniversityDepartment of Bioengineering, Faculty of Chemical and Metallurgical Engineering, Yildiz Technical UniversityDepartment of Bioengineering, Faculty of Chemical and Metallurgical Engineering, Yildiz Technical UniversityAbstract Toxoplasmosis is caused by T. gondii and can create serious health problems in humans and also worldwide economic harm. Because of the clinical and veterinary importance of toxoplasmosis, its timely and accurate diagnosis has a major impact on disease-fighting strategies. T. gondii surface antigen 1 (SAG1), an immunodominant-specific antigen, is often used as a diagnostic tool. Therefore, the aim of this study was the optimization of novel fluorescein isothiocyanate (FITC) labeling of the SAG1-specific IgY antibody to show the potential for immunofluorescence imaging of T. gondii in vitro. The specificity of IgY antibodies was controlled by an enzyme-linked immunosorbent assay (ELISA), and the concentration of the IgY antibody was detected using a spectrophotometer. The optimum incubation time and FITC concentration were determined with a fluorescence spectrometer. The obtained FITC-labeled IgY was used for marking T. gondii tachyzoites, which were cultured in vitro and viewed using light microscopy. The interaction of the fluorescence-labeled antibody and the T. gondii tachyzoites was examined with a fluorescence microscope. In this study, for the first time, a FITC-labeled anti-SAG1 IgY antibody was developed according to ELISA, fluorescence spectrometer, and fluorescence imaging of cell culture. In the future, the obtained FITC-labeled T. gondii tachyzoites’ specific IgY antibodies may be used as diagnostic tools for the detection of T. gondii infections in different samples.https://doi.org/10.1038/s41598-017-00930-1
collection DOAJ
language English
format Article
sources DOAJ
author Mehtap Sert
Rabia Cakir Koc
Yasemin Budama Kilinc
spellingShingle Mehtap Sert
Rabia Cakir Koc
Yasemin Budama Kilinc
Novel Fitc-Labeled Igy Antibody: Fluorescence Imaging Toxoplasma Gondii In Vitro
Scientific Reports
author_facet Mehtap Sert
Rabia Cakir Koc
Yasemin Budama Kilinc
author_sort Mehtap Sert
title Novel Fitc-Labeled Igy Antibody: Fluorescence Imaging Toxoplasma Gondii In Vitro
title_short Novel Fitc-Labeled Igy Antibody: Fluorescence Imaging Toxoplasma Gondii In Vitro
title_full Novel Fitc-Labeled Igy Antibody: Fluorescence Imaging Toxoplasma Gondii In Vitro
title_fullStr Novel Fitc-Labeled Igy Antibody: Fluorescence Imaging Toxoplasma Gondii In Vitro
title_full_unstemmed Novel Fitc-Labeled Igy Antibody: Fluorescence Imaging Toxoplasma Gondii In Vitro
title_sort novel fitc-labeled igy antibody: fluorescence imaging toxoplasma gondii in vitro
publisher Nature Publishing Group
series Scientific Reports
issn 2045-2322
publishDate 2017-04-01
description Abstract Toxoplasmosis is caused by T. gondii and can create serious health problems in humans and also worldwide economic harm. Because of the clinical and veterinary importance of toxoplasmosis, its timely and accurate diagnosis has a major impact on disease-fighting strategies. T. gondii surface antigen 1 (SAG1), an immunodominant-specific antigen, is often used as a diagnostic tool. Therefore, the aim of this study was the optimization of novel fluorescein isothiocyanate (FITC) labeling of the SAG1-specific IgY antibody to show the potential for immunofluorescence imaging of T. gondii in vitro. The specificity of IgY antibodies was controlled by an enzyme-linked immunosorbent assay (ELISA), and the concentration of the IgY antibody was detected using a spectrophotometer. The optimum incubation time and FITC concentration were determined with a fluorescence spectrometer. The obtained FITC-labeled IgY was used for marking T. gondii tachyzoites, which were cultured in vitro and viewed using light microscopy. The interaction of the fluorescence-labeled antibody and the T. gondii tachyzoites was examined with a fluorescence microscope. In this study, for the first time, a FITC-labeled anti-SAG1 IgY antibody was developed according to ELISA, fluorescence spectrometer, and fluorescence imaging of cell culture. In the future, the obtained FITC-labeled T. gondii tachyzoites’ specific IgY antibodies may be used as diagnostic tools for the detection of T. gondii infections in different samples.
url https://doi.org/10.1038/s41598-017-00930-1
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AT yaseminbudamakilinc novelfitclabeledigyantibodyfluorescenceimagingtoxoplasmagondiiinvitro
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