Abstract
We constructed biotinylated fusion proteins that linked to three detection tags to GAD65 at the N-terminus, and expressed them in an E. coli expression system. The Biotin14-GAD65 protein exhibited the strongest binding to both the GAD65 antibody and the streptavidin among the three constructs. We describe the optimal conditions using a Biotin14-GAD65-based immunoassay for the detection of GAD65 antibody.
Publication types
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Evaluation Study
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Validation Study
MeSH terms
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Antibodies / analysis*
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Antibodies / immunology
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Antigen-Antibody Complex / analysis
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Biosensing Techniques / methods*
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Biotin / chemistry*
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Biotin / genetics*
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Biotin / metabolism
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Enzyme-Linked Immunosorbent Assay / methods*
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Glutamate Decarboxylase / chemistry*
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Glutamate Decarboxylase / genetics*
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Glutamate Decarboxylase / metabolism
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Isoenzymes / chemistry*
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Isoenzymes / genetics*
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Isoenzymes / metabolism
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Protein Binding
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Protein Engineering / methods*
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Radioimmunoassay
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Recombinant Fusion Proteins
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Reproducibility of Results
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Sensitivity and Specificity
Substances
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Antibodies
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Antigen-Antibody Complex
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Isoenzymes
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Recombinant Fusion Proteins
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Biotin
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Glutamate Decarboxylase
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glutamate decarboxylase 2