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Somatostatin receptor staining in FFPE sections using a ligand derivative dye as an alternative to immunostaining

The confirmation of target expression in tissues is a prerequisite for molecular-targeted therapy. However, difficulties are sometimes associated with the production of appropriate antibodies against receptors. We herein developed a ligand derivative dye for the staining of receptors. The somatostat...

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Autores principales: Hasegawa, Koki, Kudoh, Shinji, Ito, Takaaki
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5300255/
https://www.ncbi.nlm.nih.gov/pubmed/28182792
http://dx.doi.org/10.1371/journal.pone.0172030
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author Hasegawa, Koki
Kudoh, Shinji
Ito, Takaaki
author_facet Hasegawa, Koki
Kudoh, Shinji
Ito, Takaaki
author_sort Hasegawa, Koki
collection PubMed
description The confirmation of target expression in tissues is a prerequisite for molecular-targeted therapy. However, difficulties are sometimes associated with the production of appropriate antibodies against receptors. We herein developed a ligand derivative dye for the staining of receptors. The somatostatin receptor (sstr) was selected as the target and FITC-octreotate as the detective agent. We performed a blot analysis to detect sstr in the transfer membrane. The sstr2 recombinant protein or cell lysate from a small cell lung carcinoma cell line (H69) was boiled and loaded onto SDS-PAGE, and the proteins were transferred to a membrane. Even after denaturing processes, FITC-octreotate still bound sstr on the membrane. Furthermore, FITC-octreotate depicted the expression of sstr in formalin-fixed and paraffin-embedded (FFPE) sections, a method that we named ligand derivative staining (LDS). The accuracies of immunostaining and LDS were compared at the points of the detection of sstr using FFPE sections of 30 neuroendocrine tumor specimens. The sensitivity of LDS was 81.8%, while those of immunostaining using anti-sstr2 and sstr5 antibodies were 72.7% and 63.6%, respectively. Thus, LDS appears to be superior to immunostaining. A ligand derivative may be used as a substitute for antibodies, and has the potential to support economical, simple, and accurate detection methods.
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spelling pubmed-53002552017-02-28 Somatostatin receptor staining in FFPE sections using a ligand derivative dye as an alternative to immunostaining Hasegawa, Koki Kudoh, Shinji Ito, Takaaki PLoS One Research Article The confirmation of target expression in tissues is a prerequisite for molecular-targeted therapy. However, difficulties are sometimes associated with the production of appropriate antibodies against receptors. We herein developed a ligand derivative dye for the staining of receptors. The somatostatin receptor (sstr) was selected as the target and FITC-octreotate as the detective agent. We performed a blot analysis to detect sstr in the transfer membrane. The sstr2 recombinant protein or cell lysate from a small cell lung carcinoma cell line (H69) was boiled and loaded onto SDS-PAGE, and the proteins were transferred to a membrane. Even after denaturing processes, FITC-octreotate still bound sstr on the membrane. Furthermore, FITC-octreotate depicted the expression of sstr in formalin-fixed and paraffin-embedded (FFPE) sections, a method that we named ligand derivative staining (LDS). The accuracies of immunostaining and LDS were compared at the points of the detection of sstr using FFPE sections of 30 neuroendocrine tumor specimens. The sensitivity of LDS was 81.8%, while those of immunostaining using anti-sstr2 and sstr5 antibodies were 72.7% and 63.6%, respectively. Thus, LDS appears to be superior to immunostaining. A ligand derivative may be used as a substitute for antibodies, and has the potential to support economical, simple, and accurate detection methods. Public Library of Science 2017-02-09 /pmc/articles/PMC5300255/ /pubmed/28182792 http://dx.doi.org/10.1371/journal.pone.0172030 Text en © 2017 Hasegawa et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Hasegawa, Koki
Kudoh, Shinji
Ito, Takaaki
Somatostatin receptor staining in FFPE sections using a ligand derivative dye as an alternative to immunostaining
title Somatostatin receptor staining in FFPE sections using a ligand derivative dye as an alternative to immunostaining
title_full Somatostatin receptor staining in FFPE sections using a ligand derivative dye as an alternative to immunostaining
title_fullStr Somatostatin receptor staining in FFPE sections using a ligand derivative dye as an alternative to immunostaining
title_full_unstemmed Somatostatin receptor staining in FFPE sections using a ligand derivative dye as an alternative to immunostaining
title_short Somatostatin receptor staining in FFPE sections using a ligand derivative dye as an alternative to immunostaining
title_sort somatostatin receptor staining in ffpe sections using a ligand derivative dye as an alternative to immunostaining
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5300255/
https://www.ncbi.nlm.nih.gov/pubmed/28182792
http://dx.doi.org/10.1371/journal.pone.0172030
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