Cargando…

A Fast Universal Immobilization of Immunoglobulin G at 4°C for the Development of Array-based Immunoassays

To maintain the antibody activity and enhance performance of array-based immunoassays, protein G was used to allow a shorter duration of immunoglobulin G immobilization at 4°C, with the antibody placed in the appropriate orientation. The multiplexed detection of six pain-related message molecules (P...

Descripción completa

Detalles Bibliográficos
Autores principales: Guo, Shu-Lin, Chen, Po-Chung, Chen, Ming-Shuo, Cheng, Yu-Che, Lin, Jun-Mu, Lee, Hoong-Chien, Chen, Chien-Sheng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3517563/
https://www.ncbi.nlm.nih.gov/pubmed/23236488
http://dx.doi.org/10.1371/journal.pone.0051370
_version_ 1782252439995416576
author Guo, Shu-Lin
Chen, Po-Chung
Chen, Ming-Shuo
Cheng, Yu-Che
Lin, Jun-Mu
Lee, Hoong-Chien
Chen, Chien-Sheng
author_facet Guo, Shu-Lin
Chen, Po-Chung
Chen, Ming-Shuo
Cheng, Yu-Che
Lin, Jun-Mu
Lee, Hoong-Chien
Chen, Chien-Sheng
author_sort Guo, Shu-Lin
collection PubMed
description To maintain the antibody activity and enhance performance of array-based immunoassays, protein G was used to allow a shorter duration of immunoglobulin G immobilization at 4°C, with the antibody placed in the appropriate orientation. The multiplexed detection of six pain-related message molecules (PRMMs) was used as examples for the development of array-based immunoassays: substance P, calcitonin gene-related peptide, nerve growth factor, brain-derived neurotrophic factor, tumor necrosis factor-α, and β-endorphin. Protein G- and non-protein G-coated slides were tested. Compared to non-protein G immunoassays, protein G shortened the antibody immobilization time at 4°C from overnight to 2 hours. Only protein G-facilitated immunoassays succeeded in simultaneously detecting all six PRMMs with high specificity. Dose-response curves showed that the limits of detection of the protein G-multiplexed immunoassays for the PRMMs was approximately 164, 167, 120, 60, 80, and 92 pg/ml, respectively. Thus, protein G effectively shortens the duration of antibody immobilization at 4°C, allowing the use of sensitive array-based immunoassays for the simultaneous detection of PRMMs.
format Online
Article
Text
id pubmed-3517563
institution National Center for Biotechnology Information
language English
publishDate 2012
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-35175632012-12-12 A Fast Universal Immobilization of Immunoglobulin G at 4°C for the Development of Array-based Immunoassays Guo, Shu-Lin Chen, Po-Chung Chen, Ming-Shuo Cheng, Yu-Che Lin, Jun-Mu Lee, Hoong-Chien Chen, Chien-Sheng PLoS One Research Article To maintain the antibody activity and enhance performance of array-based immunoassays, protein G was used to allow a shorter duration of immunoglobulin G immobilization at 4°C, with the antibody placed in the appropriate orientation. The multiplexed detection of six pain-related message molecules (PRMMs) was used as examples for the development of array-based immunoassays: substance P, calcitonin gene-related peptide, nerve growth factor, brain-derived neurotrophic factor, tumor necrosis factor-α, and β-endorphin. Protein G- and non-protein G-coated slides were tested. Compared to non-protein G immunoassays, protein G shortened the antibody immobilization time at 4°C from overnight to 2 hours. Only protein G-facilitated immunoassays succeeded in simultaneously detecting all six PRMMs with high specificity. Dose-response curves showed that the limits of detection of the protein G-multiplexed immunoassays for the PRMMs was approximately 164, 167, 120, 60, 80, and 92 pg/ml, respectively. Thus, protein G effectively shortens the duration of antibody immobilization at 4°C, allowing the use of sensitive array-based immunoassays for the simultaneous detection of PRMMs. Public Library of Science 2012-12-07 /pmc/articles/PMC3517563/ /pubmed/23236488 http://dx.doi.org/10.1371/journal.pone.0051370 Text en © 2012 Guo 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, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Guo, Shu-Lin
Chen, Po-Chung
Chen, Ming-Shuo
Cheng, Yu-Che
Lin, Jun-Mu
Lee, Hoong-Chien
Chen, Chien-Sheng
A Fast Universal Immobilization of Immunoglobulin G at 4°C for the Development of Array-based Immunoassays
title A Fast Universal Immobilization of Immunoglobulin G at 4°C for the Development of Array-based Immunoassays
title_full A Fast Universal Immobilization of Immunoglobulin G at 4°C for the Development of Array-based Immunoassays
title_fullStr A Fast Universal Immobilization of Immunoglobulin G at 4°C for the Development of Array-based Immunoassays
title_full_unstemmed A Fast Universal Immobilization of Immunoglobulin G at 4°C for the Development of Array-based Immunoassays
title_short A Fast Universal Immobilization of Immunoglobulin G at 4°C for the Development of Array-based Immunoassays
title_sort fast universal immobilization of immunoglobulin g at 4°c for the development of array-based immunoassays
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3517563/
https://www.ncbi.nlm.nih.gov/pubmed/23236488
http://dx.doi.org/10.1371/journal.pone.0051370
work_keys_str_mv AT guoshulin afastuniversalimmobilizationofimmunoglobulingat4cforthedevelopmentofarraybasedimmunoassays
AT chenpochung afastuniversalimmobilizationofimmunoglobulingat4cforthedevelopmentofarraybasedimmunoassays
AT chenmingshuo afastuniversalimmobilizationofimmunoglobulingat4cforthedevelopmentofarraybasedimmunoassays
AT chengyuche afastuniversalimmobilizationofimmunoglobulingat4cforthedevelopmentofarraybasedimmunoassays
AT linjunmu afastuniversalimmobilizationofimmunoglobulingat4cforthedevelopmentofarraybasedimmunoassays
AT leehoongchien afastuniversalimmobilizationofimmunoglobulingat4cforthedevelopmentofarraybasedimmunoassays
AT chenchiensheng afastuniversalimmobilizationofimmunoglobulingat4cforthedevelopmentofarraybasedimmunoassays
AT guoshulin fastuniversalimmobilizationofimmunoglobulingat4cforthedevelopmentofarraybasedimmunoassays
AT chenpochung fastuniversalimmobilizationofimmunoglobulingat4cforthedevelopmentofarraybasedimmunoassays
AT chenmingshuo fastuniversalimmobilizationofimmunoglobulingat4cforthedevelopmentofarraybasedimmunoassays
AT chengyuche fastuniversalimmobilizationofimmunoglobulingat4cforthedevelopmentofarraybasedimmunoassays
AT linjunmu fastuniversalimmobilizationofimmunoglobulingat4cforthedevelopmentofarraybasedimmunoassays
AT leehoongchien fastuniversalimmobilizationofimmunoglobulingat4cforthedevelopmentofarraybasedimmunoassays
AT chenchiensheng fastuniversalimmobilizationofimmunoglobulingat4cforthedevelopmentofarraybasedimmunoassays