Evaluation of poly- and monoclonal antibody-based sandwich enzyme-linked immunosorbent assay (ELISA) for their performance to detect crustacean residues in processed foods

Understanding the Mechanism of Increased IgG/IgE Reactivity but Decreased Immunodetection Recovery in Thermally Induced Shrimp (Litopenaeus vannamei) Tropomyosin via Multispectroscopic and Molecular Dynamics Simulation Techniques

“…To comply with labeling guidelines and safeguard the health of food-allergic consumers, sELISA is most commonly used as a simple, high-throughput, low-cost, highly sensitive, and specific technique for analyzing allergens in foods. , Heat treatments resulted in a considerably lower recovery of TM according to sELISA analysis, and the maximal underestimation recovery was observed in autoclaved TM with 39.2 and 74.8% reduction in PcAb sELISA tests and 50.1 and 91.6% reduction in mAb sELISA tests when compared to raw TM (Figure ). Similarly, a significant reduction in TM recovery (48.48–90.44%) also was observed in the investigation of various cooked shrimps (Litopenaeus vannamei) .…”

“…Protein marker, bovine serum albumin (BSA), isopropy-β-d-thiogalactoside (IPTG), and Tween 20 were purchased from Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China). Rat and rabbit antishrimp TM IgG and antishrimp TM monoclonal antibody (mAb) were obtained as described in our previous assay, with titers of 2.56 × 10 6 , 1.28 × 10 6 , and 4.096 × 10 6 in rat antisera, rabbit antisera, and mAb, respectively. Horseradish peroxidase (HRP)-labeled rabbit-anti-rat IgG and goat-anti-rabbit IgG were obtained from Bai Aotong Experimental Materials Center (Luoyang, Henan Province, China).…”

“…Sandwich ELISA Analysis. The sELISA analysis was carried out following the method previously reported by Zhao et al 9 In brief, plates were first coated with mice anti-TM sera (diluted 1:100 000 with CBS) and blocked with 1% (w/v) BSA, as described above. In the assay, TM standards or differently treated samples with equal concentration (100 μL/well) were added and incubated for 20 min at 37 °C.…”

“…Accurate food labeling information is urgently needed so that allergy sufferers can assess the potential health risks of food products and avoid them on a dietary basis . Additionally, as the group of the “big eight” foods, crustaceans are required to be declared and highlighted on food packaging to alert the consumer of their presence in a food product by the Codex Alimentarius (Food Code), United States (Food Allergen Labelling & Consumer Protection Act of 2004, FALCPA), and European Union (Regulation EU1169/2011). , To detect crustacean residues, although numerous highly sensitive methods have been developed, such as enzyme-linked immunosorbent assay (ELISA), , polymerase chain reaction (PCR) assay, , and liquid chromatography–mass spectrometry (LC–MS), , a protein-based immunorecognition method, the ELISA is the most heavily used technique for allergen quantification because of its simplicity, high-throughput detection capabilities, low cost, high sensitivity, and specificity. , In ELISA tests, specific and high-affinity antibodies are applied to detect biomarkers present in finished food that indicate the presence of offending ingredients. However, although stable allergens are frequently used as targets for immunodetection, unlike native protein targets, one of the major drawbacks of ELISA is the poorer recovery in detecting denatured proteins since they undergo structural changes during processing. , …”

A comprehensive overview of emerging processing techniques and detection methods for seafood allergens

“…Recently, J. Zhao et al. (2022) also developed a poly‐ and monoclonal antibody‐based (PcAb and mAb) sandwich ELISA method to detect crustacean tropomyosin in foods, and this novel method showed better matrix‐tolerance and detectability in processed foods.…”