Dataset Information

Protein-bound polyphenols create "ghost" band artifacts during chemiluminescence-based antigen detection.

ABSTRACT: Antigen detection during Western blotting commonly utilizes a horseradish peroxidase-coupled secondary antibody and enhanced chemiluminescent substrate. We utilized this technique to examine the impact of green tea-derived polyphenols on the binding of egg white protein-specific IgE antibodies from allergic human plasma to their cognate antigens. Our experiments unexpectedly showed that green tea-derived polyphenols, when stably complexed with egg white proteins, caused "ghost" band formation in the presence of horseradish peroxide. This study suggests that caution should be taken when evaluating polyphenol-bound proteins by enhanced chemiluminescence Western blotting using horseradish peroxidase and demonstrates that protein-bound polyphenols can be a source of "ghost" band artifacts on Western blots.

SUBMITTER: Plundrich N

PROVIDER: S-EPMC5497812 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Protein-bound polyphenols create "ghost" band artifacts during chemiluminescence-based antigen detection.

Plundrich Nathalie N Lila Mary Ann MA Foegeding Edward E Laster Scott S

F1000Research 20170313

Antigen detection during Western blotting commonly utilizes a horseradish peroxidase-coupled secondary antibody and enhanced chemiluminescent substrate. We utilized this technique to examine the impact of green tea-derived polyphenols on the binding of egg white protein-specific IgE antibodies from allergic human plasma to their cognate antigens. Our experiments unexpectedly showed that green tea-derived polyphenols, when stably complexed with egg white proteins, caused "ghost" band formation in ...[more]

PMID: 28721200

Similar Datasets

Project description:BackgroundProcalcitonin (PCT) is an acute phase response protein, which can be used as an indicator for early diagnosis of infection. At present, the main detection methods for PCT are electrochemiluminescence and enzyme-linked immunofluorescence. We aimed to explore the accuracy of PCT determination in a domestic chemiluminescence detection system and its correlation with other systems.MethodsClinical specimens were collected, and the precision, linearity, biological reference interval, contamination rate, Clinical reportable scope, and methodological comparison of the determination of PCT in a Chinese chemiluminescence detection system were evaluated and preliminarily verified by referring to Clinical and Laboratory Standards Institute (CLSI) documents or industry standards.ResultsThe results of precision verification showed that the coefficient of variation (CV) values of the variation coefficient of precision in the samples of low and high values were 2.07% and 0.83% respectively, while the CV values of the total variation coefficient of precision were 3.05% and 1.81% respectively; these findings all met the experimental requirements. The results of linear verification test showed that the linear range was 0.006-96.96 ng/mL, and the linear relationship was well within the detection range (R2 =0.9891). The biological reference interval and the carrying contamination rate were also verified. The clinical reportable range was 0.02-369.585 ng/mL. The results showed that the correlation coefficient between the Mindray CL900I and the Roche E602 was 0.9986, and that between the Mindray CL900I and the Snibe 2000 was 0.983. Meanwhile, when the PCT was higher than 0.1 ng/mL, the correlation coefficient was 100%.ConclusionsThe domestic chemiluminescence detection system has a good performance in the determination of calcitonin, as indicated by the measures of precision, linearity, biological reference interval, carrying contamination rate, and Clinical reportable scope, and can thus be used for clinical specimen detection. The results of methodological comparison showed that the correlation coefficient between the Mindray CL900I and Roche E602 was 0.9986, while the correlation coefficient between the Mindray CL900I and the Snibe 2000 was 0.983. The test results were consistent with the experimental requirements.

Dataset Information

Protein-bound polyphenols create "ghost" band artifacts during chemiluminescence-based antigen detection.

Publications

Protein-bound polyphenols create "ghost" band artifacts during chemiluminescence-based antigen detection.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets