Journal
FOODS
Volume 10, Issue 8, Pages -Publisher
MDPI
DOI: 10.3390/foods10081708
Keywords
ELISA; antibody; non-specific binding; cross-reaction; validation
Categories
Funding
- National Institute of Food and Agriculture, U.S. Department of Agriculture [2017-70001-25984]
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A study on the development of ELISA for food safety and quality control highlighted the critical factors leading to false immunodetection results and emphasized the importance of antibody and assay validation.
Different types of enzyme-linked immunosorbent assays (ELISA) have been widely used to control food safety and quality. To develop an accurate and reproducible ELISA, false immunodetection results caused by non-specific binding (NSB) and cross-reaction must be prevented. During the case study of sandwich ELISA development for the detection of porcine hemoglobin (P-Hb), several critical factors leading to NSB and cross-reaction were found. First, to reduce the NSB of the target analyte, the selection of microplate and blocker was discussed. Second, cross-reactions between enzyme-labeled secondary antibodies and sample proteins were demonstrated. In addition, the function of (3-aminopropyl)triethoxysilane (APTES) was evaluated. Overall, this study highlights the essence of both antibody and assay validation to minimize any false-positive/negative immunodetection results.
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