Gluten-related Diseases Research Articles

Gluten-Free Muffin: Effect of Composite Flours (Rice, Pumpkin and Unripe Banana) on The Physicochemical Properties and Sensory Attributes

Muffins are popular breakfast or snack items and are enjoyed in various cultures around the world. It is typically sweet and can come in various flavours such as blueberry, chocolate chip or banana nut. It is one among numerous baked items frequently crafted using wheat flour. However, individuals with gluten-related diseases such as celiac disorder and gluten intolerance are unable to consume wheat flour. In this research, gluten-free muffins made with a composite flour of rice, pumpkin and unripe banana with different ratios F1 (1:1:1), F2 (3:2:1), F3 (2:1:3) and F4 (1:3:2) were applied. The gluten-free muffins prepared with F3 (2 rice: 1 pumpkin: 3 unripe banana) composite flour showed the highest values of springiness and chewiness among all the samples. The gluten-free muffins using F2 (3 rice: 2 pumpkin: 1 unripe banana) composite flours were the highest for lightness and b* yellowness values. In addition, these muffins also obtained the highest percentages for ash and crude fiber contents. The 9-point scale hedonic test showed that consumers' preferences for gluten-free muffins with a ratio of 2 rice: 1 pumpkin: 3 unripe banana flours (F3) were the highest in scores as compared to the other gluten-free muffins. Thus, the study suggested that F3 which contains 2 rice: 1 pumpkin: 3 unripe banana flours was selected as the best formulation to make gluten-free muffins.

Differential pattern of neurotoxicity induced by the gliadin peptides p31-43 and p57-68 in in vitro model of epilepsy

Epilepsy is a central nervous system (CNS) disorder causing repeated seizures due to a transient excessive or synchronous alteration in the electrical activity of the brain. Several neurological disorders have been associated to gluten-related diseases (GRD), including epilepsy. However, the molecular mechanisms that associate GRD and epileptogenesis are still unknown. Our previous data have shown that the gliadin peptide 31–43 (p31-43) enhanced number and duration of seizures induced by kainate in mice and exacerbated CA3-kainate-induced neurotoxicity in organotypic hippocampal slices. Here, we investigated whether another important gliadin peptide p57-68 may exerts effects similar to p31-43 on kainate-induced neurotoxicity. We find that both peptides exacerbate kainate-induced damage in the CA3 region once simultaneously challenged. However, after pre-incubation, p31-43 additionally exacerbates neurotoxicity in the CA1 region, while p57-68 does not. These data suggested differential intracellular mechanisms activated by the peptides. Indeed, analysing intracellular signalling pathways we discover that p31-43 induces significant intracellular changes, including increased phosphorylation of Akt, Erk1/2, and p65, decreased p38 phosphorylation, and deacetylation of nuclear histone-3. Based on these observations, we demonstrate that p31-43 likely activates specific intracellular signaling pathways involved in neuronal excitability, inflammation, and epigenetic regulation, which may contribute to its exacerbation of kainate-induced neurotoxicity. In contrast, p57-68 appears to exert its effects through different mechanisms. Further research is necessary to elucidate the precise mechanisms by which these peptides influence neurotoxicity and understand their implications for neurological disorders.

Reducing Immunoreactivity of Gluten Peptides by Probiotic Lactic Acid Bacteria for Dietary Management of Gluten-Related Diseases.

Immunoreactive gluten peptides that are not digested by peptidases produced by humans can trigger celiac disease, allergy and non-celiac gluten hypersensitivity. The aim of this study was to evaluate the ability of selected probiotic strains to hydrolyze immunoreactive gliadin peptides and to identify peptidase-encoding genes in the genomes of the most efficient strains. Residual gliadin immunoreactivity was measured after one- or two-step hydrolysis using commercial enzymes and bacterial peptidase preparations by G12 and R5 immunoenzymatic assays. Peptidase preparations from Lacticaseibacillus casei LC130, Lacticaseibacillus paracasei LPC100 and Streptococcus thermophilus ST250 strains significantly reduced the immunoreactivity of gliadin peptides, including 33-mer, and this effect was markedly higher when a mixture of these strains was used. In silico genome analyses of L. casei LC130 and L. paracasei LPC100 revealed the presence of genes encoding peptidases with the potential to hydrolyze bonds in proline-rich peptides. This suggests that L. casei LC130, L. paracasei LPC100 and S. thermophilus ST250, especially when used as a mixture, have the ability to hydrolyze immunoreactive gliadin peptides and could be administered to patients on a restricted gluten-free diet to help treat gluten-related diseases.

Gluten related disorders: a review of the literature

Gluten-related disorders have become an epidemiologically significant phenomenon, with an estimated prevalence of about 5% worldwide. The current classification of gluten-related disorders is based mainly on their pathophysiology, and the following categories can be distinguished: Immune-mediated disorders, which include celiac disease, allergic reactions, such as wheat allerg, and non-celiac gluten sensitivity, a condition characterized by both gastrointestinal and extraintestinal symptoms, which has recently gained popularity. Diagnostic errors in gluten-associated diseases are still a pressing clinical problem. Based on the literature, we propose a practical methodological approach to guide clinicians on how to recognize them and avoid errors. The first part of the review presents literature data on the history, causal factors in the development of gluten-related diseases, and autoimmune gluten intolerance celiac disease.

Features of the intestinal microbiome in patients with gluten-sensitive diseases who are on a gluten-free diet

Background. Intestinal dysbiosis is associated with violating both quantitative and qualitative intestinal microbiome (IM) composition. It accompanies different gastrointestinal disorders, including non-celiac gluten sensitivity (NCGS) and celiac disease (CD). A gluten-free diet is the only existing treatment for CD and NCGS. One of the questions of interest is the characteristics of the IM of patients with gluten-related diseases and its relationship with diet therapy. Materials and methods. The study included 25 adults, 14 (56 %) with CD and 11 (44 %) with NCGS; all were on agliadin diet. We determined the faecal content of the Bacteroidetes, Firmicutes, and Actinobacteria and the rest bacterial DNA (“other” indicator) using the real-time polymerase chain reaction. Results. The Bacteroidetes content was 28.4 (Q1-Q3 9.195–37.83) % in CD and 24.98 (Q1-Q3 9.615–30.597) % in NCGS patients, p = 0.565. Firmicutes content was 53.47 (Q1-Q3 49.98–56.798) % in CD patients and 53.0 (Q1-Q3 48.12–68.53) % in NCGS, p = 0.763. The Actinobacteria content was 4.8 (Q1-Q3 3.82–6.84) % in CD patients and 5.37 (Q1-Q3 4.09–7.77) % in NCGS, p = 0.572. There was a moderate positive correlation with the diet duration (p = 0.397, p = 0.03). The Firmicutes/Bacteroidetes ratio was 1.996 (Q1-Q3 1.35–6.85) in CD patients and 2.0 (Q1-Q3 1.65–7.58) in NCGS patients, p = 0.681. The content of “other” types of IM was 15.39 (Q1-Q3 7.44–22.33) % in CD patients and 13.78 (Q1-Q3 10.65–15.58) % in NCGS patients, p = 0.936. Conclusions. We found similar intestinal microbiome features in patients with celiac disease and non-celiac gluten sensitivity. The detected intestinal microbiome changes are most likely to be a consequence of dietary features in such patients, namely the side effect of maintaining an agliadin diet.

Development of a new recombinant antibody, selected by phage-display technology from a celiac patient library, for detection of gluten in foods

Gluten, a group of ethanol-soluble proteins present in the endosperm of cereals, is extensively used in the food industry due to its ability to improve dough properties. However, gluten is also associated with a range of gluten-related diseases (GRDs), such as wheat allergies, celiac disease, and gluten intolerance. The recommended treatment for GRDs patients is a gluten-free diet. To monitor adherence to this diet, it is necessary to develop gluten-detection systems in food products. Among the available methods, immunodetection systems are the most popular due to their simplicity, reproducibility, and accuracy. The aim of this study was to generate novel high-affinity antibodies against gluten to be used as the primary reactant in an enzyme-linked immunosorbent assay (ELISA) test. These antibodies were developed by constructing an immune library from mRNA obtained from two celiac patients with a high humoral response to gluten-related proteins. The resulting library (composed by 1.1x107) was subjected to selection against gliadin using phage display technology. Following several rounds of selection, the Fab-C was selected, and demonstrated good functionality in ELISA tests, presenting a limit of detection of 15 mg/kg for detection of gluten in spiked mixtures and food products. The methodology can discriminate gluten-free products according to the current legislation.

Нецелиакийная чувствительность к глютену – диагностическая проблема в педиатрии

The possibility of a causal relationship between gluten intake and the development of symptoms in the absence of celiac disease (CD) and wheat allergy (WA) has recently gained new scientific interest and includes a condition known as non-celiac gluten sensitivity (NCGS). To date, NCGS has been widely described in adults, whereas there is little information about this pathology in pediatric practice. This article presents a clinical case of NCGS in a 6-year-old girl who was admitted to the gastroenterology department of the Republican Specialized Scientific and Practical Medical Center of Pediatrics (Republic of Uzbekistan) with complaints of abdominal pain, bloating, weight loss, liquid stools, headaches, and fatigue. The complaints appeared 5 months before hospitalization. Antibodies to tissue transglutaminase IgA were within reference ranges, specific IgE antibodies to wheat and gluten were not elevated. A biopsy from the distal duodenum revealed Marsh-Oberhuber 1 lesions. The patient was switched to a gluten-free diet (GFD) with suspected NCGS. At week 4 of the diet, an improvement was noted, but abdominal bloating persisted. An intolerance to components such as fermentable oligosaccharides, disaccharides, monosaccharides, and polyols (FODMAP) was suspected. The use of the FODMAP diet significantly improved the condition. The reintroduction of gluten after 6 weeks resulted in a recurrence of symptoms, confirming the diagnosis of NCGS. Given that the number of gluten-related diseases is increasing, and not much is known about NCGS, we decided to present this case to raise awareness and questions regarding diagnosis, the need for specific monitoring, and treatment. Key words: FODMAP, gluten, children, non-celiac gluten sensitivity, celiac disease

Characterization of α-gliadin alleles of Japanese wheat cultivars in relation to flour dough extensibility and celiac disease epitopes

Alpha-gliadins are major seed storage proteins of wheat and are involved in dough extensibility, which is related to the end-use properties of flour. However, α-gliadin causes gluten-related diseases such as celiac disease (CD). To characterize the α-gliadin allele types of Japanese cultivars, PCR markers that detect the presence of each α-gliadin gene in Chinese Spring wheat were used to determine allele-types. We surveyed 95 accessions of the Japanese wheat core collection (JWC) and 22 commercial cultivars and found 2, 10, and 4 allele types at the loci Gli-A2, Gli-B2, and Gli-D2, respectively. According to the allele-type frequency, the α-gliadin alleles of JWC showed diversities comparable to those of foreign cultivars. Allele diversities were compared within four populations of JWC, classified by genome-wide single nucleotide polymorphisms. The degree of diversity of alleles reflected the population structures, except for high diversity of the allele-types at Gli-B2 in the classical varieties. The frequencies of alleles related to end-use were higher in newer varieties than in classical varieties and landraces. Allele types that dramatically decreased and increased CD epitopes were not found in JWC, suggesting that selection(s) of α-gliadin genes was not linked to CD epitopes in Japanese cultivars.

Proteins from Modern and Ancient Wheat Cultivars: Impact on Immune Cells of Healthy Individuals and Patients with NCGS.

In non-celiac gluten sensitivity (NCGS), the elimination of wheat results in a clear symptom improvement, but gluten has still not been proven as (the sole) trigger. Due to the increase in the prevalence of gluten-related diseases, the breeding of high-performance wheat cultivars is discussed as a trigger. To analyze the immune stimulation and signal pathways, the immune cells of healthy subjects and patients with NCGS were stimulated with gliadins from wheat, and the expression and secretion of interleukin 1ß (IL1ß) and interleukin 6 (IL6) were studied. To determine the impact of wheat breeding, the monocyte cell line THP1 and human immune cells were stimulated with gliadin, glutenin, and albumin/globulin fractions of ancient and modern cereals, and expression of inflammatory molecules was checked. Immune cells of patients with NCGS showed an increased expression of IL1ß and IL6 after stimulation with gliadins compared to immune cells of healthy controls. Gliadins caused a strong activation of P-STAT3 in immune cells of healthy controls, and inhibitors of JAK and NFκB pathways considerably reduced this response. In addition to gliadins, we further showed that glutenins and albumin/globulins from all wheat cultivars from the last century, and especially from einkorn and spelt, also markedly induced the expression of inflammatory genes in THP1 and human immune cells. There was no correlation between enhanced immune stimulation and ancient or modern cultivars. This does not support the hypothesis that modern wheat breeding is responsible for the increase in gluten-related diseases. An altered immune situation is suggested in patients with NCGS.

Purified Clinoptilolite-Tuff as an Efficient Sorbent for Gluten Derived from Food.

Various gluten-related diseases (celiac disease, wheat allergy, gluten sensitivity) are known and their incidence is growing. Gluten is a specific type of plant storage protein that can impair the health of gluten-prone persons following consumption, depending on the origin. The most severe effects are induced by wheat, barley, and rye. The only treatment is based on the absolute avoidance of those foods, as even traces might have severe effects on human well-being. With the goal of binding gluten impurities after ingestion, an in vitro setting was created. A special processed kind of zeolite, purified clinoptilolite-tuff (PCT), was implemented as an adsorber of gluten derived from different origins. Zeolites are known for their excellent sorption capacities and their applications in humans and animals have been studied for a long time. Tests were also performed in artificial gastric and intestinal fluids, and the adsorption capacity was determined via a certified validated method (ELISA). Depending on the kind of gluten source, 80–130 µg/mg of gluten were bound onto PCT. Hence, purified clinoptilolite-tuff, which was successfully tested for wheat, barley, and rye, proved to be suitable for the adsorption of gluten originating from different kinds of crops. This result might form the basis for an expedient human study in the future.

Chemical and physical properties of canna (Canna edulis) and jack bean (Canavalia ensiformis)-based composite flours

Poor dietary patterns and unhealthy lifestyles are common risk factors for noncommunicable ailments, the leading causes of global death rates. Therefore, improvement of diet quality is necessary and a gluten-free diet would be the right option. This diet is not solely for patients with gluten-related diseases but has also become a trend for consumers, due to the health benefits of gluten avoidance. Underutilized food crops, including tubers and legumes, are potential raw materials for producing gluten-free food. In this study, the physicochemical properties of canna (Canna edulis) and jack bean (Canavalia ensiformis) -based composite flours were investigated. The composite flours were produced using canna flour to jack bean flour ratios of F1 (85:15), F2 (70:30), and F3 (55:45). Based on the experiment’s result, an increase in jack bean flour proportion significantly (p<0.05) increased the composite flour’s protein content, lightness, and resistant starch content of composite flours, but significantly (p<0.05) reduced carbohydrate content, oil holding capacity, and swelling power. According to the scoring test result, F2 (70:30) has the highest result and was selected as the best formula. Thus, a combination of canna and jack bean flours indicated promising properties of gluten-free flour for functional food alternative resources.

Diagnostic evaluation of microscopic enteritis in duodenal biopsies of suspected malabsorption cases with clinico-immunohistological correlation

Microscopic Enteritis (ME) is characterized by increase in intraepithelial lymphocytes (IEL) in intestinal mucosa. It represents a common feature of broad group of diseases including gluten mediated and non gluten related diseases. Duodenal biopsies play an important role in diagnosing these group of disorders. To compare IEL counts in Hematoxylin and eosin (H & E) stain and CD3 immunohistochemical (IHC) stain in duodenal biopsies of suspected malabsorption cases and compare them with clinical, immunological and biochemical parameters. This was a prospective study of two years. 164 patients were studied. IEL counted at villous tip and base in H&E sections and IHC were compared in duodenal biopsy. Data of clinical history, other parameters were collected and correlated whenever available. To compare any two variables, Chi - square test and independent T test was used. Statistical significance was defined as p < 0.05. Out of 164 cases,105 cases had increased IEL. The age range was 4 to 94 years with mean age of 43.29 ± 17.96 years. Males (56, 53.3%) were affected more than females (49, 46.6%). The clinical and histological parameters showing statistical significance with raised IEL were pallor, dyspepsia, loss of appetite, crypt architecture, blunting, ulcer, villous crypt ratio, exudates, reactive atypia and edema with P< 0.05. IEL at villous tip in CD3 had highest sensitivity (100%) and specificity (92.20%) in our study.ME should be investigated and diagnosed in correlation with a detail clinical history, complete haematological, biochemical and serological findings.

Assessment of Consumer Knowledge of Gluten-Free Food

The gluten-free food market has become one of the fastest-growing food markets for food intolerances. People with celiac disease, gluten intolerance, non-celiac gluten sensitivity, and other gluten-related diseases should follow a gluten-free diet. The demand for gluten-free products is growing yearly, mainly due to the increasing number of diagnosed cases. The aim of the study was to assess consumers’ knowledge of gluten-free foods and covered people of all ages, and was conducted using a questionnaire. The research results indicate that most consumers have a lot of knowledge concerning gluten-free food. Moreover, most consumers know what gluten is and what products to avoid while on a gluten-free diet. They also know which diseases it causes and that it is necessary to follow a diet, but they have difficulty determining the acceptable gluten content in gluten-free products. Consumers’ most crucial parameter when choosing a gluten-free product is taste. The results of the survey also show that a gluten-free diet is more popular among women than men.

Overlap-синдром целіакії з атиповим аутизмом у дитини: клінічний випадок

У статті наведені дані літературних джерел і власне спостереження overlap-синдрому целіакії з атиповим аутизмом у дитини. Розглянуто питання етіопатогенетичного зв’язку глютензалежних захворювань і розладів аутистичного спектра. Проаналізовано особливості клінічного фенотипу у випадку поєднання шлунково-кишкових проявів целіакії та характерних для розладів аутистичного спектра симптомів. Акцентовано увагу на труднощах інтерпретації та оцінки гастроінтестинальних симптомів у хворих з психічними розладами. Обговорено послідовність проведення скринінгових тестів для виключення целіакії у хворих з розладами аутистичного спектра за наявності або відсутності гастроінтестинальної симптоматики.

Multiple Wheat Genomes Reveal Novel Gli-2 Sublocus Location and Variation of Celiac Disease Epitopes in Duplicated α-Gliadin Genes.

The seed protein α-gliadin is a major component of wheat flour and causes gluten-related diseases. However, due to the complexity of this multigene family with a genome structure composed of dozens of copies derived from tandem and genome duplications, little was known about the variation between accessions, and thus little effort has been made to explicitly target α-gliadin for bread wheat breeding. Here, we analyzed genomic variation in α-gliadins across 11 recently published chromosome-scale assemblies of hexaploid wheat, with validation using long-read data. We unexpectedly found that the Gli-B2 locus is not a single contiguous locus but is composed of two subloci, suggesting the possibility of recombination between the two during breeding. We confirmed that the number of immunogenic epitopes among 11 accessions varied. The D subgenome of a European spelt line also contained epitopes, in agreement with its hybridization history. Evolutionary analysis identified amino acid sites under diversifying selection, suggesting their functional importance. The analysis opens the way for improved grain quality and safety through wheat breeding.

The effect of nitrogen fertilization on the expression of protein in wheat and tritordeum varieties using a proteomic approach

Wheat, an essential ingredient for several bakery preparations, is also responsible for gluten-related diseases in sensitive subjects. The effect of the N fertilization rate (80 vs 160 kg N ha−1) on gluten protein expression profile has been evaluated considering two soft wheats (landrace and modern) and one tritordeum cultivar (cv), grown in the same experimental field in North Italy. The proteins of refined flour were characterized through advanced proteomic approaches, including chromatography (RP-HPLC) and electrophoresis. A static model system was used to simulate in vitro digestion and the digestome peptides were examined by mass spectrometry and in silico approaches, to investigate the celiac and allergenic sequences. The CD-toxic epitopes in the digested samples were quantified by means of a R5 ELISA assay.The N fertilization rate increased the grain protein content, but it did not lead to any difference in gluten composition, with exception of glu/glia ratio in the modern wheat cv. Moreover, the gluten composition and the occurrence of toxic/allergenic epitopes varied to a great extent, according mostly to the genotype. A lower immunoreactivity, determined using R5 ELISA, was detected for the digested tritordeum flours than for the landrace (-51%) or modern (-58%) cvs, while no significant difference was observed for the N rates between each genotype. In silico analysis showed that tritordeum has fewer CD epitopes belonging to the ω-gliadins and a lower LMW-GS than the landrace or modern cv. Tritordeum presented fewer α-gliadin allergenic epitopes than the modern wheat cv. The lower frequency of celiac epitopes in tritordeum, compared to the old and the modern wheat, is probably due to the absence of a D genome.

Emergence of Celiac disease and Gluten-related disorders in Asia.

Celiac disease (CeD) is a systemic, immune-mediated enteropathy, which is triggered by gluten protein in genetically susceptible individuals. CeD, once thought to be an uncommon disease, is now recognized to affect approximately 40-60 million people globally. While CeD is now well reported from a few Asian countries such as India, China, Pakistan, and Middle Eastern countries; it is still believed to be uncommon in the rest of Asia. Gluten-related diseases other than CeD, like non-celiac gluten sensitivity (NCGS) are also emerging globally. CeD and NCGS may present with either intestinal or extra-intestinal symptoms, and a proportion of them have overlapping symptoms with irritable bowel syndrome. Hence, many of them are misdiagnosed as having irritable bowel syndrome in clinical practice. In this review, we discuss the emergence of CeD and other gluten-related disorders, both globally and in Asia, the overlapping manifestations between gluten-related disorders and irritable bowel syndrome, and the challenges associated with diagnosis and management of CeD in Asia.

Celiac disease, wheat allergy, and nonceliac sensitivity to gluten: topical issues of the pathogenesis and diagnosis of gluten-associated diseases

Historically, wheat and other gluten-containing grains have been an essential part of the diet of many people worldwide. However, the widespread use of gluten-containing products has led to an increased incidence of adverse reactions associated with their consumption, such as celiac disease and wheat allergy, as well as nonceliac gluten sensitivity, which has become common in recent years.
 Gluten-associated diseases have similar clinical manifestations (abdominal pain, bloating, diarrhea, nausea, and vomiting). The pathogenetic mechanisms underlying celiac disease and wheat allergy are quite well understood; in both cases, an immune response occurs when wheat proteins are consumed, both with effective treatment. Nonceliac gluten sensitivity is the subject of discussion; however, the processes underlying this disease are not fully understood; thus, its diagnosis and treatment have no unified approach. To date, nonceliac gluten sensitivity is a diagnosis of exclusion, which is established in the absence of markers of celiac disease or wheat allergy and improved following a gluten-free diet.
 A gluten-free diet is the most effective treatment for gluten-related diseases. However, like any other strict dietary restriction, gluten avoidance can result in reduced intakes of important nutrients, such as dietary fiber, protein, and micronutrients. In recent years, an increasing trend is found in the general population without confirmed gluten-related disorders that gluten-free product consumption or gluten-free diet adherence since gluten avoidance can improve well-being or gluten can be toxic for all human beings
 According to current guidelines, only patients diagnosed with celiac disease or wheat allergy are advised to follow a strict gluten-free diet.
 Herein, the modern conception of the immunopathology of gluten-related diseases and an overview of new potential therapies are presented.

The Gluten Gene: Unlocking the Understanding of Gluten Sensitivity and Intolerance.

Wheat flour is one of the most important food ingredients containing several essential nutrients including proteins. Gluten is one of the major protein components of wheat consisted of glutenin (encoded on chromosome 1) and gliadin (encoded on chromosome 1 and 6) and there are around hundred genes encoding it in wheat. Gluten proteins have the ability of eliciting the pathogenic immune responses and hypersensitivity reactions in susceptible individuals called “gluten-related disorders (GRDs)”, which include celiac disease (CD), wheat allergy (WA), and non-celiac gluten sensitivity (NCGS). Currently removing gluten from the diet is the only effective treatment for mentioned GRDs and studies for the appropriate and alternative therapeutic approaches are ongoing. Accordingly, several genetic studies have focused on breeding wheat with low immunological properties through gene editing methods. The present review considers genetic characteristics of gluten protein components, focusing on their role in the incidence of gluten-related diseases, and genetic modifications conducted to produce wheat with less immunological properties.

From Polyclonal Sera to Recombinant Antibodies: A Review of Immunological Detection of Gluten in Foodstuff

Gluten is the ethanol-soluble protein fraction of cereal endosperms like wheat, rye, and barley. It is widely used in the food industry because of the physical–chemical properties it gives to dough. Nevertheless, there are some gluten-related diseases that are presenting increasing prevalences, e.g., celiac disease, for which a strict gluten-free diet is the best treatment. Due to this situation, gluten labeling legislation has been developed in several countries around the world. This article reviews the gluten immune detection systems that have been applied to comply with such regulations. These systems have followed the development of antibody biotechnology, which comprise three major methodologies: polyclonal antibodies, monoclonal antibodies (mAbs) derived from hybridoma cells (some examples are 401.21, R5, G12, and α-20 antibodies), and the most recent methodology of recombinant antibodies. Initially, the main objective was the consecution of new high-affinity antibodies, resulting in low detection and quantification limits that are mainly achieved with the R5 mAb (the gold standard for gluten detection). Increasing knowledge about the causes of gluten-related diseases has increased the complexity of research in this field, with current efforts not only focusing on the development of more specific and sensitive systems for gluten but also the detection of protein motifs related to pathogenicity. New tools based on recombinant antibodies will provide adequate safety and traceability methodologies to meet the increasing market demand for gluten-free products.