Recombinant food allergens

Lorenz, Anne-Regine; Scheurer, Stephan; Haustein, Dieter; Vieths, Stefan

doi:10.1016/s0378-4347(01)00086-x

Cited by 36 publications

(21 citation statements)

References 177 publications

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“…We have suggested that the measurement of specific IgE to -5 gliadin and HMW-glutenin instead of gluten is useful to diagnose WDEIA (13). Recently, recombinant food allergens have been produced and tried to apply for diagnosis in many food allergies (25,26). Recombinant proteins of -5 gliadin and HMW-glutenin have not been available in enough quality and quantity.…”

Section: Discussionmentioning

confidence: 99%

Specific IgE Determination to Epitope Peptides of ω-5 Gliadin and High Molecular Weight Glutenin Subunit Is a Useful Tool for Diagnosis of Wheat-Dependent Exercise-Induced Anaphylaxis

Matsuo

Kohno

Niihara

et al. 2005

The Journal of Immunology

165

126

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Wheat ω-5 gliadin and a high m.w. glutenin subunit (HMW-glutenin) have been reported as major allergens in wheat-dependent exercise-induced anaphylaxis. A simultaneous detection of specific IgE to epitope sequences of both proteins is considered to be a reliable method for diagnosis of wheat-dependent exercise-induced anaphylaxis. However, the IgE-binding epitope of HMW-glutenin remains unknown. The aim of this study was to determine the IgE-binding epitopes of HMW-glutenin to establish a useful system of identifying patients with wheat-dependent exercise-induced anaphylaxis. For determination of IgE-binding epitopes of HMW-glutenin overlapping peptides were synthesized and reactivities of IgE Abs in the sera of patients to those peptides were analyzed. Three IgE-binding epitopes, QQPGQ, QQPGQGQQ, and QQSGQGQ, were identified within primary sequence of HMW-glutenin. Epitope peptides, which include IgE-binding sequences of ω-5 gliadin and a HMW-glutenin, were synthesized and peptide-specific IgE Abs were measured by CAP-System fluorescent enzyme immunoassay. Twenty-nine of 30 patients with wheat-dependent exercise-induced anaphylaxis had specific IgE Abs to these epitope peptides. None of the 25 sera from healthy subjects reacted to both epitope peptides. Twenty-five patients with atopic dermatitis who had specific IgE to wheat and/or gluten had very low or nonexistent levels of epitope peptide-specific IgE Abs. These results indicated that measurement of IgE levels specific to epitope peptides of ω-5 gliadin and HMW-glutenin is useful as an in vitro diagnostic method for the assessment of patients with wheat-dependent exercise-induced anaphylaxis.

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Section: Discussionmentioning

confidence: 99%

Specific IgE Determination to Epitope Peptides of ω-5 Gliadin and High Molecular Weight Glutenin Subunit Is a Useful Tool for Diagnosis of Wheat-Dependent Exercise-Induced Anaphylaxis

Matsuo

Kohno

Niihara

et al. 2005

The Journal of Immunology

165

126

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“…There is growing evidence that the use of panels of recombinant allergens, termed component-resolved diagnostics [3], instead of crude extracts from the allergenic sources, provides a refined molecular analysis of sensitization patterns, thus enhancing diagnosis, prediction of cross-sensitizations and strategies of immunotherapy [1][2][3]. However, a previous step for the use of recombinant allergens in diagnostic panels is to prove that their immunological and physicochemical properties are equivalent to those of their natural counterparts [4,5].…”

Section: Introductionmentioning

confidence: 99%

Recombinant lipid transfer protein Tri a 14: a novel heat and proteolytic resistant tool for the diagnosis of baker's asthma

Palacín¹,

Varela

Quirce

et al. 2009

Clin Experimental Allergy

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Summary Background Baker's asthma is an important occupational allergic disease. Wheat lipid transfer protein (LTP) Tri a 14 is a major allergen associated with wheat allergy. No panel of wheat recombinant allergens for component‐resolved diagnosis of baker's asthma is currently available. Objective To evaluate the potential role of recombinant Tri a 14 as a novel tool for the diagnosis of baker's asthma, and to test the heat and proteolytic resistance of the wheat LTP allergen. Methods A cDNA encoding Tri a 14 was isolated and sequenced, the recombinant allergen produced in Pichia pastoris and purified by chromatographic methods. Physicochemical and immunological comparison of the natural and recombinant forms of Tri a 14 was carried out by N‐terminal amino acid sequencing, matrix‐assisted laser desorption/ionization mass spectrometry, circular dichroism (CD) analysis, IgE immunodetection, and specific IgE determination and ELISA‐inhibition assays using a pool or individual sera from 26 patients with baker's asthma. Thermal denaturation and simulated gastrointestinal digestion of both Tri a 14 forms were checked by spectroscopic and electrophoretic methods, respectively, and biological activity by basophil activation test (BAT). Results Natural and recombinant Tri a 14 were similarly folded, as indicated by their nearly identical CD spectra and heat denaturation profiles. A high interclass correlation coefficient (0.882) was found between specific IgE levels to both Tri a 14 proteins in individual sera from baker's asthma patients, but a slightly lower IgE‐binding potency of rTri a 14 was detected by ELISA‐inhibition assays. Natural and recombinant Tri a 14 elicited positive BAT in two and one out of three patients, respectively. Heat denaturation profiles and simulated gastrointestinal digestion assays indicated that Tri a 14 displayed a high heat and digestive proteolytic resistance, comparable to those of peach Pru p 3, the model food allergen of the LTP family. Conclusions Recombinant Tri a 14 is a potential tool for baker's asthma diagnosis, based on its physicochemical and immunological similarity with its natural counterpart. Wheat Tri a 14 shows a high thermal stability and resistance to gastrointestinal digestion.

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“…This may be due to the differences of recognization of tertiary structure of DIII and DIII* under ELISA, since both type of specific IgE antibodies (against sequential and tertiary epitopes) can be recognized by ELISA. As pointed by Lorenz and Vieths (14), recombinant food allergens are, in general, not identical copies of the natural analogue, since changes to the IgE binding properties of the recombinant protein may occur. The allergenic reactivity may be altered or even be eliminated through Note.…”

Section: Resultsmentioning

confidence: 99%

“…Recombinant proteins can be produced in large quantities and with great purity, making it easier to study the molecular design of the protein when attempting to reduce the allergenic structure by genetic engineering. To date, more than 40 food allergens from fruits, vegetables, nuts, milk, and seafood have been expressed as recombinant proteins (14). To study the structurefunction relationship of DIII allergenicity, the recombinant ovomucoid third domain (DIII*) was produced using an Escherichia coli expression system.…”

mentioning

confidence: 99%

IgE Binding Properties of the Recombinant Ovomucoid Third Domain Expressed in Escherichia coli

Sasaki

Mine

2001

Biochemical and Biophysical Research Communications

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Recombinant food allergens

Cited by 36 publications

References 177 publications

Specific IgE Determination to Epitope Peptides of ω-5 Gliadin and High Molecular Weight Glutenin Subunit Is a Useful Tool for Diagnosis of Wheat-Dependent Exercise-Induced Anaphylaxis

Specific IgE Determination to Epitope Peptides of ω-5 Gliadin and High Molecular Weight Glutenin Subunit Is a Useful Tool for Diagnosis of Wheat-Dependent Exercise-Induced Anaphylaxis

Recombinant lipid transfer protein Tri a 14: a novel heat and proteolytic resistant tool for the diagnosis of baker's asthma

IgE Binding Properties of the Recombinant Ovomucoid Third Domain Expressed in Escherichia coli

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