Development of monoclonal antibody-based ELISA for the quantification of orange allergen Cit s 2 in fresh and processed oranges

Kiyota, Kyohei; Kawatsu, Kentaro; Sakata, Junko; Yoshimitsu, Masato; Akutsu, Kazuhiko; Satsuki-Murakami, Taro; Ki, Masami; Kajimura, K.; Yamano, Tetsuo

doi:10.1016/j.foodchem.2017.03.140

Cited by 15 publications

(10 citation statements)

References 23 publications

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“…The limit of detection was 31.3 ng/mL, with a coefficient of variation (CV) of less than 30%, and the quantification range was determined to be from 62.5 to 500 ng/mL, with a CV of less than 10% in 11 experiments, as previously reported (Kiyota et al, 2017).…”

Section: Parameters Of Indirect Elisa Based On the Anti-sola L 1 Peptsupporting

confidence: 68%

Detection of the tomato allergen Sola l 1 and evaluation of its reactivity after heat and papain treatment

Kiyota

Yoshimitsu

Satsuki-Murakami

et al. 2017

Food and Agricultural Immunology

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Section: Parameters Of Indirect Elisa Based On the Anti-sola L 1 Peptsupporting

confidence: 68%

Detection of the tomato allergen Sola l 1 and evaluation of its reactivity after heat and papain treatment

Kiyota

Yoshimitsu

Satsuki-Murakami

et al. 2017

Food and Agricultural Immunology

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“…The absence of detected Man i1 in these two samples might have been due to their containing too little Man i1 to be detected. Kiyota et al (2017) analyzed the food products containing orange allergen (Cit s 2) by sELISA based monoclonal antibody, and only two of 11 samples were detected (Kiyota et al, 2017). The orange allergen was not detected because it was below the LOD.…”

Section: Resultsmentioning

confidence: 99%

Development of monoclonal antibody‐based sandwich ELISA for detecting major mango allergen Man i1 in processed foods

Tsai

Yin

Chen

et al. 2021

Journal of Food Safety

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Mango (Mangifera indica L.) is a popular tropical fruit but also an allergenic source. In Taiwan, the packaging of any food product that contains mango must display an allergen warning. Therefore, establishing a reliable method for detecting mango residues in food products is important. In this study, a sandwich enzyme‐linked immunosorbent assay (sELISA) was developed to detect the major mango allergen Man i1. First, five batches of polyclonal and monoclonal antibodies against Man i1 were generated and used to identify the optimal antibody pair for sELISA. Using 2G5 mAb as the capture antibody and anti‐rMan i1 pAb as the detection antibody yielded the strongest assay signal. The limit of detection (LOD) and limit of quantification (LOQ) of the developed ELISA are estimated to be 3.9 and 21.2 ng/ml rMan i1, respectively, after optimization. The assay exhibits good specificity to mango and offers good precision and reproducibility since the calculated coefficient of variation (CV) of the intra‐ and inter‐assays were 5.5–11.7% and 7.4–12.4%, respectively. Finally, 23 commercial food products were evaluated using the developed sELISA and the results of this assay closely agreed (95.7%) with those of the Western blot. Thus, the developed sELISA provides a potential method for detecting mango allergens in foods for protecting people with mango allergy from the accidental ingestion of mango.

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“…To prepare the antibody for detecting orange allergen Cit s 2, we decided to produce the recombinant protein of Cit s 2 as an immunoantigen. The expression of recombinant Cit s 2 (rCit s 2)‐small ubiquitin‐like modifier (SUMO) was performed as described previously (Kiyota et al, ). Briefly, based on the information from a public database (United States Department of Energy), the Cit s 2‐coding region was amplified using a polymerase chain reaction (PCR) using cDNA as a template and the following primer pairs: 5′‐ATG TCG TGG CAA GCT TAC GTC‐3′ and 5′‐CTA AAG ACC CTG ATC AAT GAG A‐3′.…”

Section: Methodsmentioning

confidence: 99%

“…Since the prepared polyclonal antibody was against a recombinant Cit s 2 protein, it was necessary to verify if it recognize the native Cit s 2 (nCit s 2). To test the nCit s 2 against the antibody, it was purified from fresh navel oranges using an affinity column as described previously (Kiyota et al, ). Briefly, the column was prepared from 7.5 g CNBr‐activated Sepharose 4B (GE Healthcare) combined with 180 mg of poly‐ l ‐proline (Sigma‐Aldrich, St. Louis, MO) with a molecular weight of 1,000–10,000.…”

Section: Methodsmentioning

confidence: 99%

Evaluation of cleaning methods for residual orange extract on different cookware materials using ELISA with profilin allergen indicator

Kiyota

Sakata

Satsuki-Murakami

et al. 2017

J Food Process Engineering

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Unintentional contamination of cookware with allergen residues causes unexpected allergic reactions to individuals with food allergies. Our aim was to find an efficient cleaning method for residual orange extract as a food allergen on cookware fabricated from four types of materials—polypropylene (PP), wood, stainless steel, and glass—after food preparation. To evaluate the cleaning methods, we developed a novel enzyme‐linked immunosorbent assay system using a rabbit polyclonal antibody against a recombinant orange profilin allergen. Using this assay system with a limit of quantification of 2,500 µg/mL, we measured the residual orange extract on the cookware surfaces using swabbing tests. Rinsing with 1 L of water showed a >95% removal efficiency for stainless steel and glass cookware, whereas half the PP and wood cookware required scrubbing with a detergent‐containing sponge for complete cleanliness. Our findings may help reduce the burden for cooks and food manufacturers in controlling fruit allergens. Practical applications The cleaning process is essential in preventing cross‐contamination of food allergens through the equipment surfaces of kitchens and food processing lines. There are no established standard cleaning processes for fruit allergens. The extracts often remain on cookware surfaces after food preparation because of their viscosity. Since even small amounts of allergens can elicit symptoms of food allergies, removing food allergens by cleaning the equipment surfaces is important to avoid unexpected food‐related allergic reactions and produce safe foods for those with food allergies. Here, we investigated efficient cleaning methods using laboratory scale tests for controlling orange allergens. We used orange extract as a representative fruit‐generated adhesive extract on different cookware materials—polypropylene, wood, stainless steel, and glass. Since these materials are commonly used in food processing equipment, our findings could help develop an efficient and economical allergy control method for both cooks and manufacturers of processed fruit without excessive cleaning.

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Development of monoclonal antibody-based ELISA for the quantification of orange allergen Cit s 2 in fresh and processed oranges

Cited by 15 publications

References 23 publications

Detection of the tomato allergen Sola l 1 and evaluation of its reactivity after heat and papain treatment

Detection of the tomato allergen Sola l 1 and evaluation of its reactivity after heat and papain treatment

Development of monoclonal antibody‐based sandwich ELISA for detecting major mango allergen Man i1 in processed foods

Evaluation of cleaning methods for residual orange extract on different cookware materials using ELISA with profilin allergen indicator

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