Anti-allergen antibodies

a technology of anti-allergen antibodies and antibodies, which is applied in the field of anti-allergen antibodies, can solve the problems of severe systemic reactions, large emergency visits, hospital admissions, and deaths each year, and achieve the effects of reducing and increasing the number of hospital admissions

Inactive Publication Date: 2020-06-25
UNIV ZURICH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0061]FIG. 6. Effect of different anti-peanut allergen antibody combinations can be evaluated by leukotriene (sLT) release assay. Leukocytes from healthy blood donors were isolated and surface IgE was removed by incubation with lactic acid. Leukocytes were re-sensitized with IgE by incubation with plasma from allergic patients. Re-sensitized leukocy

Problems solved by technology

Systemic anaphylaxis, as manifested by urticaria, angioedema, bronchospasm, diarrhea, dysrhythmias, and cardiovascular collapse, is responsible for a large number of emergency visits, hospital admissions, and deaths each year.
Indeed, allergy to peanuts is the most frequent cause of food-related death.
Individuals may be so sensitive to peanut allergens that severe systemic reactions can occur in response to minute contaminants of the allergen int

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

of Peanut-Secific Antibodies by ELISA Assay

[0245]Enzyme linked immunosorbent assay (ELISA) was used to evaluate allergen reactivity in the sera of allergic patients. Allergens (Ara h 2, Ara h 1, Ara h 3, Ara h 6, peanut extract) were coated on plates and anti-peanut antibody levels in sera from allergic patients were detected using HRP-conjugated anti-human antibody. Altogether, sera from 84 allergic patients, were used in the assays. The following protocol describes the experimental procedures for the detection of anti-allergen antibodies by ELISA assay. Allergic patients have shown seroreactivity against several Ara h peanut allergens, implicating the suitability of these antibodies against allergic response induced by several peanut allergens.

[0246]ELISA for the Detection of Peanut-Specific Antibodies

[0247]96 well microplates (Costar®, Corning Incorporated, Corning, N.Y., USA) were coated with peanut allergens either extracted and purified from peanuts or recombinantly expressed....

example 2

A Determination of the Antibodies of the Present Invention

[0248]EC50 binding of exemplary anti-peanut antibodies of the present invention to peanut allergens or peanut extract, was determined by ELISA. Serial dilutions of MAbs (from 1000 ng / ml down to 0.0169 ng / ml) were incubated for 2 hours with antigen-coated plates (coating overnight at 4° C. or 1 h at 37° C. with 1 μg / ml antigen in PBS, followed by wash out and blocking with 2% BSA in PBS). The plates were subsequently washed and binding of MAbs was detected with anti-human HRP-conjugated Fc-gamma-specific secondary antibody (Jackson ImmunoResearch, West Grove, Pa., USA). Concentrations of MAb resulting in half of maximal binding to respective antigens (EC50, ng / ml) were calculated using GraphPad Prism 5 software on sigmoidal dose-response curves (variable slope, 4 parameters) obtained by plotting the log of the concentration versus OD450 nm measurements; for the results see FIG. 3 and table 2.

TABLE 2EC50 ELISA determination of ...

example 3

petition with Human-Mouse Chimeric Constructs of Anti-Peanut Antibodies

[0249]As first step of mapping, differential binding of anti-peanut MAbs of the present invention to distinct antigen binding sites was examined to determine the number of different binding sites. For this purpose, MAbs were expressed either with human (hMAb) or mouse (hmMab) Fc and cross-competition experiments were carried out by coating antigen on plates and by detecting binding of human Mabs in the presence of titrated amount of hmMabs. Detection of hMAbs bound to the ligand was performed by a HRP conjugated secondary antibody directed against the Fc portion of the primary antibody (Jackson ImmunoResearch, West Grove, Pa., USA). As may be seen from Table 3, exemplary anti-peanut allergens antibodies 37D5 and 12G10 of the present invention compete each other for binding of Ara h 2 but not with antibodies 4B2 and 32B10, indicating that 12G10 and 37D5 bind other sites of Ara h 2 than 4B2 and 32B10.

TABLE 3The dif...

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Abstract

The present invention generally relates to antibodies or binding fragments thereof capable of binding an allergen, in particular a food allergen as well as pharmaceutical compositions comprising such antibodies or binding fragments thereof for the treatment of allergy, in particular food allergy. In addition the invention relates to methods for evaluating the capacity of a candidate antibody or binding fragment thereof to inhibit allergen binding/and/or allergen- induced activity in a human and methods of detecting or quantifying whether an allergen is present in a sample.

Description

FIELD OF THE INVENTION[0001]The present invention generally relates to antibodies or binding fragments thereof capable of binding an allergen, in particular a food allergen as well as pharmaceutical compositions comprising such antibodies or binding fragments thereof for the treatment of allergy, in particular food allergy. In addition the invention relates to methods for evaluating the capacity of a candidate antibody or binding fragment thereof to inhibit allergen binding / and / or allergen-induced activity in a human and methods of detecting or quantifying whether an allergen is present in a sample.BACKGROUND OF THE INVENTION[0002]Food allergies are reactions to ingested foods that may lead to clinical manifestations from skin, respiratory and gastrointestinal symptoms up to severe and life-threatening reactions, i.e. systemic anaphylaxis. Allergic reactions to many different types of foods have been described, but some of the most common are reactions to peanuts and shellfish. Such...

Claims

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Application Information

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IPC IPC(8): C07K16/16A61P37/08
CPCC07K2317/21C07K2317/34A61P37/08A61K31/137C07K16/16A61K2039/577
Inventor AGUZZI, ADRIANOWUILLEMIN, NATASCHASONATI, TIZIANABIELI, DIMITRI
Owner UNIV ZURICH
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