Ultrasensitive monoclonal antibodies specific to thermal stable-soluble proteins of buckwheat

Kim, Sol-A; Toushik, Sazzad Hossen; Lee, Jeong Eun; Shim, Won‐Bo

doi:10.1016/j.foodchem.2023.136269

Cited by 4 publications

(5 citation statements)

References 30 publications

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“…Notably, the total soluble protein concentration of the processed fat and meat extracts by hot extraction was much higher than that of the extracts treated with cold extraction. The hot extraction may have increased the protein extraction efficiency because the fat and meat tissue are expanded by heat to form a space, and the extraction buffer penetrates this space to increase the contact area ( Kim et al, 2023 ). This result means that hot extraction is more effective than cold extraction for extracting TSSP to be analyzed from processed fat and meat.…”

Section: Resultsmentioning

confidence: 99%

“…After performing the iELISA, absorbance was measured at 405 nm, and each condition showing the highest sensitivity was chosen as an optimal condition for the iELISA. Additionally, the test sample exhibiting an absorbance of 0.2 less, corresponding to the absorbance of the negative sample+5 SD of the absorbance of the negative sample, was judged to be negative in iELISA ( Kim et al, 2023 ).…”

Section: Methodsmentioning

confidence: 99%

“…Additionally, the test sample exhibiting an absorbance of 0.2 less, corresponding to the absorbance of the negative sample + 5 standard deviations (SD) of the absorbance of the negative sample, was judged to be negative in iELISA (Kim et al, 2023).…”

Section: Optimization Of An Ielisa For the Analysis Of Tssp In Pork F...mentioning

confidence: 99%

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A Highly Sensitive Indirect Enzyme-Linked Immunosorbent Assay (ELISA) Based on a Monoclonal Antibody Specific to Thermal Stable-Soluble Protein in Pork Fat for the Rapid Detection of Pork Fat Adulterated in Heat-Processed Beef Meatballs

Kim,

Lee,

Kim

et al. 2023

Food Sci Anim Resour

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Processed foods containing pork fat tissue to improve flavor and gain economic benefit may cause severe issues for Muslims, Jews, and vegetarians. This study aimed to develop an indirect enzyme-linked immunosorbent assay (iELISA) based on a monoclonal antibody specific to thermal stable-soluble protein in pork fat tissue and apply it to detect pork fat tissue in heat-processed (autoclave, steam, roast, and fry) beef meatballs. To develop a sensitive iELISA, the optimal sample pre-cooking time, coating conditions, primary and secondary dilution time, and various buffer systems were tested. The change in the iELISA sensitivity with different 96-well microtiter microplates was confirmed. The detection limit of iELISA performed with an appropriate microplate was 0.015% (w/w) pork fat in raw and heat-treated beef. No cross-reactions to other meats or fats were shown. These results mean that the iELISA can be used as an analytical method to detect trace amounts of pork fat mixed in beef.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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A Highly Sensitive Indirect Enzyme-Linked Immunosorbent Assay (ELISA) Based on a Monoclonal Antibody Specific to Thermal Stable-Soluble Protein in Pork Fat for the Rapid Detection of Pork Fat Adulterated in Heat-Processed Beef Meatballs

Kim,

Lee,

Kim

et al. 2023

Food Sci Anim Resour

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“…In the indirect ELISA analysis, a sample showing more than 0.2 OD value (OD value of 0% peanut sample + 5 × SD of the OD value of 0% peanut sample) was classified to be a positive sample [ 22 ]. As shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Goat anti-mouse IgG (H+L) peroxidase conjugate diluted (1:10,000) was added with 100 μl each well and incubated at 37°C for 1 h. After adding ABTS solution to develop color, the results of the indirect ELISA were measured at 405 nm using a microplate reader. The limit of detection (LOD) was obtained using the method described by Kim et al (2023) [22].…”

Section: Optimization Of Indirect Elisa For Peanut Detectionmentioning

confidence: 99%

Detection of a Thermal Stable-Soluble Protein (TSSP) as a Marker of Peanut Adulteration Using a Highly Sensitive Indirect Enzyme-Linked Immunosorbent Assay based on Monoclonal Antibodies

Kim

Toushik

Lee

et al. 2023

J. Microbiol. Biotechnol.

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Food allergy represents a severe problem for many societies, including sensitive populations, academies, health authorities, and the food industry. Peanut allergy occupies a special place in the food allergy spectrum. To prevent consumption by consumers suffering from a peanut allergy, a rapid and sensitive detection method is essential to identify unintended peanut adulteration in processed foods. In this study, we produced four monoclonal antibodies (MAbs; RO 3A1-12, PB 4C12-10, PB 5F9-23, and PB 6G4-30) specific to thermo-stable and soluble proteins (TSSPs) of peanut and developed an enzyme-linked immunosorbent assay (ELISA) based on the MAbs. Among them, PB 5F9-23 MAb was firmly bound to Ara h 1, and other MAbs strongly reacted to Ara h 3 in the Western blot analysis. An antibody cocktail solution of the MAbs was used to enhance the sensitivity of an indirect ELISA, and the limit of detection of the indirect ELISA based on the antibody cocktail solution was 1 ng/ml and improved compared to the indirect ELISA based on the single MAb (11 ng/ml). The cross-reaction analysis revealed the high specificity of developed MAbs to peanut TSSPs without cross-reaction to other food allergens, including nuts. Subsequently, analyzing processed foods by indirect ELISA, all foods labeled as containing peanuts in the product description were confirmed to be positive. The results indicate that the developed antibodies exhibit high specificity and sensitivity to peanuts and can be used as bio-receptors in immunoassays or biosensors to detect intentional or unintentional adulteration of peanuts in processed foods, particularly heat-processed foods.

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Colorimetric and surface-enhanced Raman scattering dual-mode lateral flow immunosensor using phage-displayed shark nanobody for the detection of crustacean allergen tropomyosin

Jiao,

Chen,

et al. 2024

Journal of Hazardous Materials

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Ultrasensitive monoclonal antibodies specific to thermal stable-soluble proteins of buckwheat

Cited by 4 publications

References 30 publications

A Highly Sensitive Indirect Enzyme-Linked Immunosorbent Assay (ELISA) Based on a Monoclonal Antibody Specific to Thermal Stable-Soluble Protein in Pork Fat for the Rapid Detection of Pork Fat Adulterated in Heat-Processed Beef Meatballs

A Highly Sensitive Indirect Enzyme-Linked Immunosorbent Assay (ELISA) Based on a Monoclonal Antibody Specific to Thermal Stable-Soluble Protein in Pork Fat for the Rapid Detection of Pork Fat Adulterated in Heat-Processed Beef Meatballs

Detection of a Thermal Stable-Soluble Protein (TSSP) as a Marker of Peanut Adulteration Using a Highly Sensitive Indirect Enzyme-Linked Immunosorbent Assay based on Monoclonal Antibodies

Colorimetric and surface-enhanced Raman scattering dual-mode lateral flow immunosensor using phage-displayed shark nanobody for the detection of crustacean allergen tropomyosin

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