Enhanced detection of single-cell-secreted proteins using a fluorescent immunoassay on the protein-G-terminated glass substrate

Yoon Jeong,1,2 Kwan Hong Lee,1,2 Hansoo Park,3 Jonghoon Choi1,2 1Department of Bionano Technology, Graduate School, Hanyang University, Seoul, 2Department of Bionano Engineering, Hanyang University ERICA, Ansan, 3School of Integrative Engineering, Chung-Ang University, Seoul, South Korea Abstract:...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Jeong Y, Lee KH, Park H, Choi J
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2015
Materias:	Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/2aede23d10ab459fba8c556d0848ad0a
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:2aede23d10ab459fba8c556d0848ad0a
record_format	dspace
spelling	oai:doaj.org-article:2aede23d10ab459fba8c556d0848ad0a2021-12-02T03:11:39ZEnhanced detection of single-cell-secreted proteins using a fluorescent immunoassay on the protein-G-terminated glass substrate1178-2013https://doaj.org/article/2aede23d10ab459fba8c556d0848ad0a2015-11-01T00:00:00Zhttps://www.dovepress.com/enhanced-detection-of-single-cell-secreted-proteins-using-a-fluorescen-peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013Yoon Jeong,1,2 Kwan Hong Lee,1,2 Hansoo Park,3 Jonghoon Choi1,2 1Department of Bionano Technology, Graduate School, Hanyang University, Seoul, 2Department of Bionano Engineering, Hanyang University ERICA, Ansan, 3School of Integrative Engineering, Chung-Ang University, Seoul, South Korea Abstract: We present an evaluation of protein-G-terminated glass slides that may contain a suitable substrate for aligning the orientation of antibodies to obtain better binding moiety to the target antigen. The results of the protein-G-terminated slides were compared with those obtained with epoxy-based slides to evaluate signal enhancement for human immunoglobulin G (IgG) targets, and an increase in the average fluorescence intensity was observed for the lowest measurable amount of IgG target in the assay using protein-G-terminated slides. Applying this strategy for signal amplification to single-cell assays improves the limits of detection for human IgG protein and cytokines (interleukin-2 and interferon-γ) captured from hybridomas. Our data indicate that protein-G-terminated slides have a higher binding capacity for antigens and have better spot-to-spot consistency than that of traditional epoxy-based slides. These properties would be beneficial in the detection of fine amounts of single-cell-secreted proteins, which may provide key insights into cell–cell communication and immune responses. Keywords: microwell array, antibody’s orientation, single cell analysis, secreted cytokine, protein-G-terminated surfaceJeong YLee KHPark HChoi JDove Medical PressarticleMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol 2015, Iss default, Pp 7197-7205 (2015)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine (General) R5-920
spellingShingle	Medicine (General) R5-920 Jeong Y Lee KH Park H Choi J Enhanced detection of single-cell-secreted proteins using a fluorescent immunoassay on the protein-G-terminated glass substrate
description	Yoon Jeong,1,2 Kwan Hong Lee,1,2 Hansoo Park,3 Jonghoon Choi1,2 1Department of Bionano Technology, Graduate School, Hanyang University, Seoul, 2Department of Bionano Engineering, Hanyang University ERICA, Ansan, 3School of Integrative Engineering, Chung-Ang University, Seoul, South Korea Abstract: We present an evaluation of protein-G-terminated glass slides that may contain a suitable substrate for aligning the orientation of antibodies to obtain better binding moiety to the target antigen. The results of the protein-G-terminated slides were compared with those obtained with epoxy-based slides to evaluate signal enhancement for human immunoglobulin G (IgG) targets, and an increase in the average fluorescence intensity was observed for the lowest measurable amount of IgG target in the assay using protein-G-terminated slides. Applying this strategy for signal amplification to single-cell assays improves the limits of detection for human IgG protein and cytokines (interleukin-2 and interferon-γ) captured from hybridomas. Our data indicate that protein-G-terminated slides have a higher binding capacity for antigens and have better spot-to-spot consistency than that of traditional epoxy-based slides. These properties would be beneficial in the detection of fine amounts of single-cell-secreted proteins, which may provide key insights into cell–cell communication and immune responses. Keywords: microwell array, antibody’s orientation, single cell analysis, secreted cytokine, protein-G-terminated surface
format	article
author	Jeong Y Lee KH Park H Choi J
author_facet	Jeong Y Lee KH Park H Choi J
author_sort	Jeong Y
title	Enhanced detection of single-cell-secreted proteins using a fluorescent immunoassay on the protein-G-terminated glass substrate
title_short	Enhanced detection of single-cell-secreted proteins using a fluorescent immunoassay on the protein-G-terminated glass substrate
title_full	Enhanced detection of single-cell-secreted proteins using a fluorescent immunoassay on the protein-G-terminated glass substrate
title_fullStr	Enhanced detection of single-cell-secreted proteins using a fluorescent immunoassay on the protein-G-terminated glass substrate
title_full_unstemmed	Enhanced detection of single-cell-secreted proteins using a fluorescent immunoassay on the protein-G-terminated glass substrate
title_sort	enhanced detection of single-cell-secreted proteins using a fluorescent immunoassay on the protein-g-terminated glass substrate
publisher	Dove Medical Press
publishDate	2015
url	https://doaj.org/article/2aede23d10ab459fba8c556d0848ad0a
work_keys_str_mv	AT jeongy enhanceddetectionofsinglecellsecretedproteinsusingafluorescentimmunoassayontheproteingterminatedglasssubstrate AT leekh enhanceddetectionofsinglecellsecretedproteinsusingafluorescentimmunoassayontheproteingterminatedglasssubstrate AT parkh enhanceddetectionofsinglecellsecretedproteinsusingafluorescentimmunoassayontheproteingterminatedglasssubstrate AT choij enhanceddetectionofsinglecellsecretedproteinsusingafluorescentimmunoassayontheproteingterminatedglasssubstrate
_version_	1718401842981896192

Enhanced detection of single-cell-secreted proteins using a fluorescent immunoassay on the protein-G-terminated glass substrate

Ejemplares similares