Novel Monoclonal Antibody-Based Immunodiagnostic Assay for Rapid Detection of Deamidated Gluten Residues

Masiri, Jongkit; Benoit, Lora; Katepalli, Madhu; Meshgi, M Aghazadeh; Cox, David L.; Nadala, Cesar; Sung, S.-S. J.; Samadpour, Mansour

doi:10.1021/acs.jafc.5b06085

Cited by 14 publications

(8 citation statements)

References 21 publications

(32 reference statements)

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“…At present, many different methods have been used to detect CIT toxin. The most commonly used methods are mass spectrometry and chromatography analysis, but these methods are unfixable for the actual sample detection because of the large equipment and professional staff necessary. , Immunoassays based on a monoclonal antibody have also been used widely in immunological detection of small molecules/toxins and pathogens in samples; ,− however, the traditional monoclonal antibodies are often costly, time-consuming, and cannot be manipulated genetically . These disadvantages severely limited its application in biological toxin detection.…”

Section: Discussionmentioning

confidence: 99%

Screening and Molecular Evolution of a Single Chain Variable Fragment Antibody (scFv) against Citreoviridin Toxin

Wang

Zhuang

et al. 2016

J. Agric. Food Chem.

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Citreoviridin (CIT), a small food-borne mycotoxin produced by Penicillium citreonigrum, is generally distributed in various cereal grains and farm crop products around the world and has caused cytotoxicity as an uncompetitive inhibitor of ATP hydrolysis. A high affinity single chain variable fragment (scFv) antibody that can detect the citreoviridin in samples is still not available; therefore, it is very urgent to prepare an antibody for CIT detection and therapy. In this study, an amplified and assembled scFv from hybridoma was used to construct the mutant phage library by error-prone PCR, generating a 2 × 10 capacity mutated phage display library. After six rounds of biopanning, the selected scFv-5A10 displayed higher affinity and specificity to CIT antigen, with an increased affinity of 13.25-fold (K = 5.7 × 10 L/mol) compared to that of the original wild-type scFv. Two critical amino acids (P100 and T151) distributed in H-CDR3 and L-FR regions that were responsible for scFv-5A10 to CIT were found and verified by oligonucleotide-directed mutagenesis, and the resulting three mutants except for the mutant (P100K) lost binding activity significantly against CIT, as predicated. Indirect competitive ELISA (ic-ELISA) indicated that the linear range to detect CIT was 25-562 ng/mL with IC at 120 ng/mL. The limit of detection was 14.7 ng/mL, and the recovery average was (90.612 ± 3.889)%. Hence, the expressed and purified anti-CIT MBP-linker-scFv can be used to detect CIT in corn and related samples.

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Section: Discussionmentioning

confidence: 99%

Screening and Molecular Evolution of a Single Chain Variable Fragment Antibody (scFv) against Citreoviridin Toxin

Wang

Zhuang

et al. 2016

J. Agric. Food Chem.

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“…(2015) produced the antibody INRA‐DG1, which can be used to detect deamidated gluten sequences (Table 1). Likewise, 2D4 monoclonal antibody was developed to recognize deamidated gliadin (Benoit, Cox, et al., 2016; Benoit, Meshgi, et al., 2016), and 2B9 monoclonal antibody‐based LFD was recently proposed as an immunodiagnostic assay for rapid detection of deamidated gluten (Masiri et al., 2016).…”

Section: Immunochemical Detection Methodsmentioning

confidence: 99%

Advances in quantification and analysis of the celiac‐related immunogenic potential of gluten

Ribeiro

Sousa

Sabença

et al. 2021

Comp Rev Food Sci Food Safe

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Gluten-free products have emerged in response to the increasing prevalence of gluten-related disorders, namely celiac disease. Therefore, the quantification of gluten in products intended for consumption by individuals who may suffer from these pathologies must be accurate and reproducible, in a way that allows their proper labeling and protects the health of consumers. Immunochemical methods have been the methods of choice for quantifying gluten, and several kits are commercially available. Nevertheless, they still face problems such as the initial extraction of gluten in complex matrices or the use of a standardized reference material to validate the results. Lately, other methodologies relying mostly on mass spectrometry-based techniques have been explored, and that may allow, in addition to quantitative analysis, the characterizationof gluten proteins. On the other hand, although the level of 20 mg/kg of gluten detected by these methods is sufficient for a product to be considered gluten-free, its immunogenic potential for celiac patients has not been clinically validated. In this sense, in vitro and in vivo models, such as the organoid technology applied in gut-on-chip devices and the transgenic humanized mouse models, respectively, are being developed for investigating both the gluten-induced pathogenesis and the treatment of celiac disease. Due to the ubiquitous nature of gluten in the food industry, as well as the increased prevalence of gluten-related disorders, here we intend to summarize the available methods for gluten quantification in food matrices and for the evaluation of its immunogenic potential concerning the development of novel therapies for celiac disease to highlight active research and discuss knowledge gaps and current challenges in this field.

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“…The G12 antibody was produced against a synthetic 33-mer (LQLQPFPQPQLPYPQPQLPYPQPQLPYPQPQPF) of α2-gliadin, believed to invoke immunopathogenicity and the antibody recognizes the QPQLPY epitope of the peptide (21, 22). Recently, a novel monoclonal antibody that recognizes deamidated gliadin was generated by Pi Bioscientific Inc. (23, 24). Detection and quantitation of intact gluten has been routinely performed using sandwich ELISAs (16–19, 21, 25–29).…”

Section: Introductionmentioning

confidence: 99%

Detection and Quantitation of Gluten in Fermented-Hydrolyzed Foods by Antibody-Based Methods: Challenges, Progress, and a Potential Path Forward

Panda

Garber

2019

Front. Nutr.

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Celiac disease (CD) affects ~1 in 141 individuals in the United States, requiring adherence to a strict gluten-free diet. The Codex Standard and the European Commission states that gluten level of gluten-free foods must not exceed 20 ppm. The FDA requires food bearing the labeling claim “gluten-free” to contain <20 ppm gluten. Accurate quantitation of gluten in fermented-hydrolyzed foods by antibody-based methods is a challenge due to the lack of appropriate reference materials and variable proteolysis. The recent uses of proteases (e.g., proline endopeptidases or PEP) to hydrolyze immunopathogenic sequences of gluten proteins further complicates the quantitation of immunopathogenic gluten. The commercially available antibody-based methods routinely used to detect and quantitate gluten are not able to distinguish between different hydrolytic patterns arising from differences in fermentation processes. This is a severe limitation that makes accurate quantitation and, ultimately, a detailed evaluation of any potential health risk associated with consuming the food difficult. Utilizing gluten-specific antibodies, a recently developed multiplex-competitive ELISA along with western blot analysis provides a potential path forward in this direction. These complimentary antibody-based technologies provide insight into the extent of proteolysis resulting from various fermentation processes and have the potential to aid in the selection of appropriate hydrolytic calibration standards, leading to accurate gluten quantitation in fermented-hydrolyzed foods.

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Novel Monoclonal Antibody-Based Immunodiagnostic Assay for Rapid Detection of Deamidated Gluten Residues

Cited by 14 publications

References 21 publications

Screening and Molecular Evolution of a Single Chain Variable Fragment Antibody (scFv) against Citreoviridin Toxin

Screening and Molecular Evolution of a Single Chain Variable Fragment Antibody (scFv) against Citreoviridin Toxin

Advances in quantification and analysis of the celiac‐related immunogenic potential of gluten

Detection and Quantitation of Gluten in Fermented-Hydrolyzed Foods by Antibody-Based Methods: Challenges, Progress, and a Potential Path Forward

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