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3 pages, 176 KiB

Open AccessEditorial

Application of Immunoassay Technology in Food Inspection

by Peipei Li and Maojun Jin

Foods 2023, 12(15), 2923; https://doi.org/10.3390/foods12152923 - 31 Jul 2023

Cited by 1 | Viewed by 1220

Abstract

Food safety is as important as ever, and the safeguards implemented to inspect and reduce pesticides, veterinary drugs, toxins, pathogens, illegal additives, and other deleterious contaminants in our food supply has helped improve human health and increase the length and quality of our [...] Read more.

Food safety is as important as ever, and the safeguards implemented to inspect and reduce pesticides, veterinary drugs, toxins, pathogens, illegal additives, and other deleterious contaminants in our food supply has helped improve human health and increase the length and quality of our lives [...] Full article

(This article belongs to the Special Issue Application of Immunoassay Technology in Food Inspection)

Research

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21 pages, 4142 KiB

Open AccessArticle

Development of New Antibodies and an ELISA System to Detect the Potato Alkaloids α-Solanine and α-Chaconine

by Kohki Okada and Kano Matsuo

Foods 2023, 12(8), 1621; https://doi.org/10.3390/foods12081621 - 12 Apr 2023

Cited by 2 | Viewed by 2450

Abstract

Food poisoning can be caused by the potato alkaloids α-solanine (SO) and α-chaconine (CHA). Therefore, this study aimed to establish new enzyme-linked immunosorbent assays (ELISAs) for detecting these two toxins in biological samples and potato extracts. Two antibodies that bind to solanidine, a [...] Read more.

Food poisoning can be caused by the potato alkaloids α-solanine (SO) and α-chaconine (CHA). Therefore, this study aimed to establish new enzyme-linked immunosorbent assays (ELISAs) for detecting these two toxins in biological samples and potato extracts. Two antibodies that bind to solanidine, a chemical compound found in both SO and CHA, were newly developed, and two types of ELISAs (Sold1 ELISA and Sold2 ELISA) were constructed. We measured SO and CHA diluted in phosphate-buffered saline (PBS), serum, and urine. The detection performance of the two ELISAs for SO and CHA in PBS was higher than in serum and urine, and the sensitivity of Sold2 ELISA was lower than that of Sold1 ELISA. Thus, we used these ELISAs to measure SO and CHA in potato part extracts and found that potato sprouts contained approximately 80-fold more SO and CHA than tubers and 8-fold more SO and CHA than peels. Although the detection sensitivity of SO and CHA depends on the sample types, these ELISAs may be effective as future clinical and food testing methods after further improvements. Full article

(This article belongs to the Special Issue Application of Immunoassay Technology in Food Inspection)

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14 pages, 5494 KiB

Open AccessArticle

Comparison of Chemiluminescence Enzyme Immunoassay (Cl-ELISA) with Colorimetric Enzyme Immunoassay (Co-ELISA) for Imidacloprid Detection in Vegetables

by Rongqi Zhai, Ge Chen, Guangyang Liu, Xiaodong Huang, Xiaomin Xu, Lingyun Li, Yanguo Zhang, Donghui Xu and A. M. Abd El-Aty

Foods 2023, 12(1), 196; https://doi.org/10.3390/foods12010196 - 1 Jan 2023

Cited by 17 | Viewed by 3208

Abstract

Imidacloprid is one of the most commonly used insecticides for managing pests, thus, improving the quality and yield of vegetables. The abuse/misuse of imidacloprid contaminates the environment and threatens human health. To reduce the risk, a colorimetric enzyme-linked immunoassay assay (Co-ELISA) and chemiluminescence [...] Read more.

Imidacloprid is one of the most commonly used insecticides for managing pests, thus, improving the quality and yield of vegetables. The abuse/misuse of imidacloprid contaminates the environment and threatens human health. To reduce the risk, a colorimetric enzyme-linked immunoassay assay (Co-ELISA) and chemiluminescence enzyme-linked immunoassay assay (Cl-ELISA) were established to detect imidacloprid residues in vegetables. The linear range of Co-ELISA ranged between 1.56 μg/L and 200 μg/L with a limit of detection (LOD) of 1.56 μg/L. The values for Cl-ELISA were 0.19 μg/L to 25 μg/L with an LOD of 0.19 μg/L, which are lower than those of Co-ELISA. Fortifying Chinese cabbage, cucumber, and zucchini with imidacloprid at 10, 50, and 100 μg/L yielded recoveries between 81.7 and 117.6% for Co-ELISA and at 5, 10, and 20 µg/L yielded recoveries range from 69.7 to 120.6% for Cl-ELISA. These results indicate that Cl-ELISA has a high sensitivity and a rapid detection time, saving cost (antigen and antibody concentrations) and serving as a more efficient model for the rapid detection of imidacloprid residue. Full article

(This article belongs to the Special Issue Application of Immunoassay Technology in Food Inspection)

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12 pages, 1803 KiB

Open AccessArticle

Enzyme-Linked Aptamer Kits for Rapid, Visual, and Sensitive Determination of Lactoferrin in Dairy Products

by Fan Zhang, Hongxia Du, Linsen Li, Tengfei Li, Jing Wang, Zilei Chen, Mengmeng Yan, Chao Zhu and Feng Qu

Foods 2022, 11(23), 3763; https://doi.org/10.3390/foods11233763 - 22 Nov 2022

Cited by 1 | Viewed by 1744

Abstract

Lactoferrin (Lf), as a popular nutritional fortification in dairy products, has the ability regulate the body’s immune system and function as a broad-spectrum antibacterial, which is of great significance to the growth and development of infants and children. Herein, an indirect competitive enzyme-linked [...] Read more.

Lactoferrin (Lf), as a popular nutritional fortification in dairy products, has the ability regulate the body’s immune system and function as a broad-spectrum antibacterial, which is of great significance to the growth and development of infants and children. Herein, an indirect competitive enzyme-linked aptamer assay (ELAA) kit was established for rapid, sensitive, and visual determination of Lf in dairy products. In the construction, the Lf aptamer was conjugated with horseradish peroxidase (HRP) as the recognition probe and aptamer complementary strand (cDNA) were anchored onto the microplate as the capture probe. The recognition probes were first mixed with a sample solution and specifically bound with the contained Lf, then added into the microplate in which the free recognition probes in the mixture were captured by the capture probe. After washing, the remaining complex of cDNA/Aptamer/HRP in the microplate was conducted with a chromogenic reaction through HRP, efficiently catalyzing the substrate 3, 3′, 5, 5′-tetramethylbenzidine (TMB), therefore the color shade would directly reflect Lf concentration. Under the optimization conditions, a good linear relationship (R², 0.9901) was obtained in the wide range of 25–500 nM with the detection limit of 14.01 nM and a good specificity, as well as the reliable recoveries. Furthermore, the ELAA kits achieved the Lf determination with an accuracy of 79.71~116.99% in eleven samples, which consisted of three kinds of dairy products: including goat milk powder, cow milk powder, and nutrition drop. Moreover, the results were also validated by the high-performance capillary electrophoresis (HPCE) method. The ELAA kit provides a simple and convenient determination for Lf in dairy products, and it is highly expected to be commercialized. Full article

(This article belongs to the Special Issue Application of Immunoassay Technology in Food Inspection)

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12 pages, 1594 KiB

Open AccessArticle

pH-Regulated Strategy and Mechanism of Antibody Orientation on Magnetic Beads for Improving Capture Performance of Staphylococcus Species

by Fuying Kang, Yin Yang, Jingwen Li, Erning Chen, Tian Hong, Lulu Zhao and Meihong Du

Foods 2022, 11(22), 3599; https://doi.org/10.3390/foods11223599 - 11 Nov 2022

Cited by 4 | Viewed by 2686

Abstract

Immunomagnetic beads (IMBs) have been widely used to capture and isolate target pathogens from complex food samples. The orientation of the antibody immobilized on the surface of magnetic beads (MBs) is closely related to the effective recognition with an antigen. We put forward [...] Read more.

Immunomagnetic beads (IMBs) have been widely used to capture and isolate target pathogens from complex food samples. The orientation of the antibody immobilized on the surface of magnetic beads (MBs) is closely related to the effective recognition with an antigen. We put forward an available strategy to orient the antibody on the surface of MBs by changing the charged amino group ratio of the reactive amino groups at optimal pH value. Quantum dots labeling antigen assay, antigen-binding fragment (Fab) accessibility assay and lysine mimicking were used for the first time to skillfully illustrate the antibody orientation mechanism. This revealed that the positively charged ε-NH₂ group of lysine on the Fc relative to the uncharged amino terminus on Fab was preferentially adsorbed on the surface of MBs with a negatively charged group at pH 8.0, resulting in antigen binding sites of antibody fully exposed. This study contributes to the understanding of the antibody orientation on the surface of MBs and the potential application of IMBs in the separation and detection of pathogenic bacteria in food samples. Full article

(This article belongs to the Special Issue Application of Immunoassay Technology in Food Inspection)

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11 pages, 2452 KiB

Open AccessArticle

An Alkyne-Mediated SERS Aptasensor for Anti-Interference Ochratoxin A Detection in Real Samples

by Hao Wang, Lu Chen, Min Li, Yongxin She, Chao Zhu and Mengmeng Yan

Foods 2022, 11(21), 3407; https://doi.org/10.3390/foods11213407 - 28 Oct 2022

Cited by 9 | Viewed by 1938

Abstract

Avoiding interference and realizing the precise detection of mycotoxins in complex food samples is still an urgent problem for surface-enhanced Raman spectroscopy (SERS) analysis technology. Herein, a highly sensitive and specific aptasensor was developed for the anti-interference detection of Ochratoxin A (OTA). In [...] Read more.

Avoiding interference and realizing the precise detection of mycotoxins in complex food samples is still an urgent problem for surface-enhanced Raman spectroscopy (SERS) analysis technology. Herein, a highly sensitive and specific aptasensor was developed for the anti-interference detection of Ochratoxin A (OTA). In this aptasensor, 4-[(Trimethylsilyl) ethynyl] aniline was employed as an anti-interference Raman reporter to prove a sharp Raman peak (1998 cm⁻¹) in silent region, which could avoid the interference of food bio-molecules in 600–1800 cm⁻¹. 4-TEAE and OTA-aptamer were assembled on Au NPs to serve as anti-interference SERS probes. Meanwhile, Fe₃O₄ NPs, linked with complementary aptamer (cApts), were applied as capture probes. The specific binding of OTA to aptamer hindered the complementary binding of aptamer and cApt, which inhibited the binding of SERS probes and capture probes. Hence, the Raman responses at 1998 cm⁻¹ were negatively correlated with the OTA level. Under the optimum condition, the aptasensor presented a linear response for OTA detection in the range of 0.1–40 nM, with low detection limits of 30 pM. In addition, the aptasensor was successfully applied to quantify OTA in soybean, grape and milk samples. Accordingly, this anti-interference aptasensor could perform specific, sensitive and precise detection of OTA in real samples, and proved a reliable reference strategy for other small-molecules detection in food samples. Full article

(This article belongs to the Special Issue Application of Immunoassay Technology in Food Inspection)

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12 pages, 2424 KiB

Open AccessArticle

Identification and Application of Two Promising Peptide Ligands for the Immunodetection of Imidacloprid Residue

by Tianyang You, Yuan Ding, Yue Huang, Yang Lu, Minghua Wang and Xiude Hua

Foods 2022, 11(20), 3163; https://doi.org/10.3390/foods11203163 - 11 Oct 2022

Cited by 2 | Viewed by 1557

Abstract

As the most widely used neonicotinoid insecticide, it is of great significance to explore the immunoreagents and immunoassays for imidacloprid (IMI) residue. In immunoassays, specific peptide ligands, such as peptidomimetic and anti-immunocomplex peptides, are regarded as promising substitutes for chemical haptens. In the [...] Read more.

As the most widely used neonicotinoid insecticide, it is of great significance to explore the immunoreagents and immunoassays for imidacloprid (IMI) residue. In immunoassays, specific peptide ligands, such as peptidomimetic and anti-immunocomplex peptides, are regarded as promising substitutes for chemical haptens. In the present work, we identified thirty sequences of peptidomimetics and two sequences of anti-immunocomplex peptides for IMI from three phage pVIII display cyclic peptide libraries, in which the anti-immunocomplex peptides are the first reported noncompetitive reagents for IMI. The peptidomimetic 1-9-H and anti-immunocomplex peptide 2-1-H that showed the best sensitivity were utilized to develop competitive and noncompetitive phage enzyme-linked immunosorbent assays (P-ELISAs), with a half inhibition concentration of 0.55 ng/mL for competitive P-ELISA and a half-saturation concentration of 0.35 ng/mL for noncompetitive P-ELISA. The anti-immunocomplex peptide was demonstrated to greatly improve the specificity compared with competitive P-ELISA. In addition, the accuracy of proposed P-ELISAs was confirmed by recovery analysis and HPLC verification in agricultural and environmental samples. These results show that the peptide ligands identified from phage display library can replace chemical haptens in the immunoassays of IMI with the satisfactory performance. Full article

(This article belongs to the Special Issue Application of Immunoassay Technology in Food Inspection)

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11 pages, 2642 KiB

Open AccessCommunication

Quantum-Dot-Bead-Based Fluorescence-Linked Immunosorbent Assay for Sensitive Detection of Cry2A Toxin in Cereals Using Nanobodies

by Yulou Qiu, Ajuan You, Xianshu Fu, Mingzhou Zhang, Haifeng Cui, Biao Zhang, Weiwei Qin, Zihong Ye and Xiaoping Yu

Foods 2022, 11(18), 2780; https://doi.org/10.3390/foods11182780 - 9 Sep 2022

Cited by 5 | Viewed by 2031

Abstract

In this study, a quantum-dot-bead (QB)-based fluorescence-linked immunosorbent assay (FLISA) using nanobodies was established for sensitive determination of the Cry2A toxin in cereal. QBs were used as the fluorescent probe and conjugated with a Cry2A polyclonal antibody. An anti-Cry2A nanobody P2 was expressed [...] Read more.

In this study, a quantum-dot-bead (QB)-based fluorescence-linked immunosorbent assay (FLISA) using nanobodies was established for sensitive determination of the Cry2A toxin in cereal. QBs were used as the fluorescent probe and conjugated with a Cry2A polyclonal antibody. An anti-Cry2A nanobody P2 was expressed and used as the capture antibody. The results revealed that the low detection limit of the developed QB-FLISA was 0.41 ng/mL, which had a 19-times higher sensitivity than the traditional colorimetric ELISA. The proposed assay exhibited a high specificity for the Cry2A toxin, and it had no evident cross-reactions with other Cry toxins. The recoveries of Cry2A from the spiked cereal sample ranged from 86.6–117.3%, with a coefficient of variation lower than 9%. Moreover, sample analysis results of the QB-FLISA and commercial ELISA kit correlated well with each other. These results indicated that the developed QB-FLISA provides a potential approach for the sensitive determination of the Cry2A toxin in cereals. Full article

(This article belongs to the Special Issue Application of Immunoassay Technology in Food Inspection)

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11 pages, 3137 KiB

Open AccessArticle

Determination of Acetamiprid Residues in Vegetables by Indirect Competitive Chemiluminescence Enzyme Immunoassay

by Zixin Zhu, Qiuyun Shi, Jianwei Wu, Kangli He, Jianguo Feng and Sa Dong

Foods 2022, 11(16), 2507; https://doi.org/10.3390/foods11162507 - 19 Aug 2022

Cited by 10 | Viewed by 1995

Abstract

Acetamiprid (ACE) is widely used in various vegetables to control pests, resulting in residues and posing a threat to human health. For the rapid detection of ACE residues in vegetables, an indirect competitive chemiluminescence enzyme immunoassay (ic-CLEIA) was established. The optimized experimental parameters [...] Read more.

Acetamiprid (ACE) is widely used in various vegetables to control pests, resulting in residues and posing a threat to human health. For the rapid detection of ACE residues in vegetables, an indirect competitive chemiluminescence enzyme immunoassay (ic-CLEIA) was established. The optimized experimental parameters were as follows: the concentrations of coating antigen (ACE-BSA) and anti-ACE monoclonal antibody were 0.4 and 0.6 µg/mL, respectively; the pre-incubation time of anti-ACE monoclonal antibody and ACE (sample) solution was 30 min; the dilution ratio of goat anti-mouse-HRP antibody was 1:2500; and the reaction time of chemiluminescence was 20 min. The half-maximum inhibition concentration (IC₅₀), the detection range (IC₁₀–IC₉₀), and the detection limit (LOD, IC₁₀) of the ic-CLEIA were 10.24, 0.70–96.31, and 0.70 ng/mL, respectively. The cross-reactivity rates of four neonicotinoid structural analogues (nitenpyram, thiacloprid, thiamethoxam, and clothianidin) were all less than 10%, showing good specificity. The average recovery rates in Chinese cabbage and cucumber were 82.7–112.2%, with the coefficient of variation (CV) lower than 9.19%, which was highly correlated with the results of high-performance liquid chromatography (HPLC). The established ic-CLEIA has the advantages of simple pretreatment and detection process, good sensitivity and accuracy, and can meet the needs of rapid screening of ACE residues in vegetables. Full article

(This article belongs to the Special Issue Application of Immunoassay Technology in Food Inspection)

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14 pages, 2724 KiB

Open AccessArticle

Design and Characterization of a Novel Hapten and Preparation of Monoclonal Antibody for Detecting Atrazine

by Lingyuan Xu, A.M. Abd El-Aty, Jae-Han Shim, Jong-Bang Eun, Xingmei Lei, Jing Zhao, Xiuyuan Zhang, Xueyan Cui, Yongxin She, Fen Jin, Lufei Zheng, Jing Wang, Maojun Jin and Bruce D. Hammock

Foods 2022, 11(12), 1726; https://doi.org/10.3390/foods11121726 - 13 Jun 2022

Cited by 6 | Viewed by 2118

Abstract

This study provides the first design and synthetic protocol for preparing highly sensitive and specific atrazine (ATR) monoclonal antibodies (mAbs). In this work, a previously unreported hapten, 2-chloro-4-ethylamino-6-isopropylamino-1,3,5-triazine, was designed and synthesized, which maximally exposed the characteristic amino group ATR to an animal [...] Read more.

This study provides the first design and synthetic protocol for preparing highly sensitive and specific atrazine (ATR) monoclonal antibodies (mAbs). In this work, a previously unreported hapten, 2-chloro-4-ethylamino-6-isopropylamino-1,3,5-triazine, was designed and synthesized, which maximally exposed the characteristic amino group ATR to an animal immune system to induce the expected antibody. The molecular weight of the ATR hapten was 259.69 Da, and its purity was 97.8%. The properties of the anti-ATR mAb were systematically characterized. One 9F5 mAb, which can detect ATR, was obtained with an IC₅₀ value (the concentration of analyte that produced 50% inhibition of ATR) of 1.678 µg/L for ATR. The molecular weight for the purified 9F5 mAb was approximately 52 kDa for the heavy chain and 15 kDa for the light chain. The anti-ATR mAb prepared in this study was the IgG₁ type. The working range of the standard curve (IC₂₀ (the concentration of analyte that produced 20% inhibition of ATR)_—IC₈₀ (the concentration of analyte that produced 80% inhibition of ATR)) was 0.384 to 11.565 µg/L. The prepared anti-ATR mAb had high specificity, sensitivity, and affinity with low cross-reactivity. The prepared anti-ATR mAb could provide the core raw material for establishing an ATR immunoassay. Full article

(This article belongs to the Special Issue Application of Immunoassay Technology in Food Inspection)

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12 pages, 2112 KiB

Open AccessArticle

Cascade-Enhanced Lateral Flow Immunoassay for Sensitive Detection of Okadaic Acid in Seawater, Fish, and Seafood

by Olga D. Hendrickson, Elena A. Zvereva, Anatoly V. Zherdev and Boris B. Dzantiev

Foods 2022, 11(12), 1691; https://doi.org/10.3390/foods11121691 - 9 Jun 2022

Cited by 16 | Viewed by 2233

Abstract

In this investigation, a new approach for developing a sensitive lateral flow immunoassay (LFIA) was proposed for the detection of the hazardous marine toxin okadaic acid (OA). It is based on the indirect format with anti-species antibodies labeled by gold nanoparticles (AuNPs) and [...] Read more.

In this investigation, a new approach for developing a sensitive lateral flow immunoassay (LFIA) was proposed for the detection of the hazardous marine toxin okadaic acid (OA). It is based on the indirect format with anti-species antibodies labeled by gold nanoparticles (AuNPs) and cascade signal amplification. The latter is performed by first passing a mixture of anti-OA antibodies and a tested sample along the immunochromatographic test strip and then performing several cycles of the interaction of anti-species antibodies conjugated with AuNPs with free antibodies, which bind to anti-species antibodies but are not specific to the target analyte. As a result, branched aggregates are formed, due to which the colorimetric signal intensification occurs. The developed test system enabled the detection of OA with an instrumental detection limit of 30 pg/mL and a cutoff of 1 ng/mL, which exceeds these characteristics in the LFIA without amplification by 7 and 2 times, respectively. The OA recoveries from seawater, fish, and seafood varied from 76.9% to 126%. The test system may be required for point-of-care monitoring of samples for phycotoxin contamination; the developed principle of signal amplification can be used in cases where highly sensitive detection of trace amounts of a contaminant is required. Full article

(This article belongs to the Special Issue Application of Immunoassay Technology in Food Inspection)

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Review

Jump to: Editorial, Research

20 pages, 358 KiB

Open AccessReview

Current Advances in Immunoassays for the Detection of β₂-Agonists

by Shuyu Ouyang, Shuting Yu and Yingying Le

Foods 2022, 11(6), 803; https://doi.org/10.3390/foods11060803 - 11 Mar 2022

Cited by 15 | Viewed by 2436

Abstract

β₂-agonists are a group of synthetic phenylethanolamine compounds which are traditionally used for treating bronchospasm. These compounds can also increase skeletal muscle mass and decrease body fat. The illegal use of β₂-agonists in food-producing animals results in residue of [...] Read more.

β₂-agonists are a group of synthetic phenylethanolamine compounds which are traditionally used for treating bronchospasm. These compounds can also increase skeletal muscle mass and decrease body fat. The illegal use of β₂-agonists in food-producing animals results in residue of β₂-agonists in edible tissues and causes adverse health effects in humans. Thus, the detection of β₂-agonists at trace level in complex sample matrices is of great importance for monitoring the abuse of β₂-agonists. Many methods have been developed to detect β₂-agonists. Among them, a variety of antigen–antibody interaction-based techniques have been established to detect β₂-agonists in various samples, including animal feed, urine, serum, milk, tissues and hair. In this review, we summarized current achievement in the extraction of β₂-agonists from testing samples and detection of β₂-agonists using immunological techniques. Future perspectives were briefly discussed. Full article

(This article belongs to the Special Issue Application of Immunoassay Technology in Food Inspection)

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