IgY antialérgenos específicos del grupo 1 de ácaros del polvo doméstico inducidos por oligopéptidos sintéticos no glicosilados

Abstract

Introduction: The use of specific antibodies capable of detecting allergens of the group 1 of house dust mites represents a potential strategy to reduce exposure and clinical symptomatology associated with asthma and allergic rhinitis. Objective: To produce and purify chicken antibodies specific for the dust mites Dermatophagoides sp. and B. tropicalis using the IgY technology. Materials and methods: We designed and synthesized oligopeptides showing immunogenic epitopes of Der p1, Der f1, and Blo t1. These were used to produce IgY antibodies in Hy Line Brown chickens. IgY were extracted from egg yolk using thiophilic chromatography. The immunogenicity and specificity were assayed by indirect ELISA and Dot Blot. Results: We obtained high reactivity of IgY antibodies against epitopes of allergens present in whole body mites extracts of D. farinae, D. pteronyssinus, and B. tropicalis. The highest IgY levels were registered between days 32 and 40 after immunization. The antibodies showed high immunoreactivity and specificity towards D. farinae proteins with detection limits above 0.03 μg of mite proteins under the experimental conditions used. Purified IgY did not show significant reactivity when binding to Periplaneta americana extract. Conclusion: The IgY technology allowed the production of specific antibodies against house dust mites group 1 allergens using non-glycosylated synthetic peptides. To our knowledge, this is the first time that this immunochemicals are used in the detection of mites of medical relevance.

Immunological detection in indoor environments of house dust mite allergens using avian antibodies - IgY: An innovative tool

“…The IgY antibodies used in the present study were polyvalent in nature, which would explain their greater reactivity. Despite the use of specific synthetic peptides for chicken immunization, IgY antibodies recognize both group-1 allergens and others present in mites' whole body extracts as reported in a previous study (Egea, et al, 2018).…”

“…Recently, our group used IgY technology to produce polyclonal antibodies against dust mite group 1 allergens (cysteine protease). These antibodies showed high reactivity and sensitivity towards D. farinae, D. pteronyssinus, and B. tropicalis allergenic proteins including group-1 mite allergens (Egea, et al, 2018;Egea, et al, 2019). Here, we used two of these IgY antibodies (IgY-PO2 and IgY-PO4) to explore their applicability for detecting mite allergens in dust samples using an indirect ELISA test.…”

“…Two polyclonal avian antibodies (IgY) from Hy Line Brown hen eggs were produced against non-glycosylated synthetic oligopeptides designed in silico based on sequences from dust mite group-1 allergens Bt (named PO2) and Dsp (named PO4) (Patent US9416163 B2) (Barrera, et al, 2016). We determined antibody titers by ELISA while in a previous study (Egea, et al, 2018), mite protein concentration and the ability of antibodies to recognize Bt and Dsp group-1 allergens had been determined by Western Blot.…”

“…We conducted an exploratory study to detect the usefulness of IgYPO2 and IgYPO4 in the detection of Bt and Dsp allergens in 100 house dust samples. Mite allergens were detected by an indirect ELISA test as previously described (Barrera, et al, 2016;Egea, et al, 2018). We coated polystyrene plates with 96 wells (Nunc Maxisorp®) with 100 µL of antigen (mite extracts or dust extracts) and incubated the plates in a wet chamber at room temperature for 18 hours.…”