TY - JOUR
T1 - A Fast Universal Immobilization of Immunoglobulin G at 4°C for the Development of Array-based Immunoassays
AU - Guo, Shu Lin
AU - Chen, Po Chung
AU - Chen, Ming Shuo
AU - Cheng, Yu Che
AU - Lin, Jun Mu
AU - Lee, Hoong Chien
AU - Chen, Chien Sheng
N1 - Copyright:
Copyright 2013 Elsevier B.V., All rights reserved.
PY - 2012/12/7
Y1 - 2012/12/7
N2 - 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.
AB - 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.
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U2 - 10.1371/journal.pone.0051370
DO - 10.1371/journal.pone.0051370
M3 - Article
C2 - 23236488
AN - SCOPUS:84870858464
VL - 7
JO - PLoS One
JF - PLoS One
SN - 1932-6203
IS - 12
M1 - e51370
ER -