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An improved assay for the diagnosis of peanut allergy

A peanut and test strip technology, used in disease diagnosis, biological testing, allergic diseases, etc., can solve problems such as inability to diagnose

Pending Publication Date: 2021-12-10
EUROIMMUN MEDIZINISCHE LABORDIAGNOSTIKA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The finding that no patients were monosensitized to Ara h 7 is another reason not to use any Ara h 7 isoform for routine diagnosis, as it is not considered to increase the sensitivity of diagnostic tests for identifying peanut-allergic patients
[0014] Currently, many patients with peanut allergies cannot be diagnosed based on the results of routine testing

Method used

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  • An improved assay for the diagnosis of peanut allergy
  • An improved assay for the diagnosis of peanut allergy
  • An improved assay for the diagnosis of peanut allergy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0131] Example 1: Detection of IgE antibodies against the Ara h 7 isotype in sera from peanut-sensitized patients by ELISA

[0132] 1.1. Recombinant protein

[0133] Heterologous expression and purification of recombinant allergens

[0134] Ara h 7.0101 (SEQ ID NO11), Ara h 7.0201 (SEQ ID NO 12), Ara h 7.0 (SEQ ID NO 13), Ara h 6 (SEQ ID NO 15) and Ara h 2 (SEQ ID NO 14) in Escherichia coli was expressed and purified as a His6 fusion protein, which has as Sitaru C, C, Probst C, Komorowski L, I, Schmidt E, Schlumberger W, Rose C, W, Zillikens D. Enzyme-linkedimmunosorbent assay using multimers of the 16th non-collagenous domain of the BP180 antigen for sensitive and specific detection of pemphigoidautoantibodies. Exp Dermatol. 2007 Sep; 16 (9): 770-7 described in the corresponding SEQ ID NO sequence shown. Purification was performed by stationary metal ion chromatography under denaturing conditions.

[0135] 1.2. Detection of IgE binding by ELISA

[0136] The IgE-bind...

Embodiment 2

[0143] Example 2: Detection of IgE Antibodies by Line Blotting

[0144] The IgE binding capacity of 2S albumin Ara h 2, Ara h 6 and three Ara h 7 isoforms was investigated by immunoblot analysis. Serum samples from 34 peanut-sensitized subjects ( figure 2 ) were incubated with the line blot-immunoblot and the intensity was assessed using EUROLINEScan software. In addition, 20 sera from atopic subjects without peanut allergy and 17 healthy blood donors were incubated as negative controls (all negative, data not shown).

[0145] For line blot immunoblotting, the recombinant allergens were coated on nylon-based membranes using standard methods. The membrane is then closed, dried and fixed on foil. The foil was cut into strips, which were incubated with serum.

[0146]Manual incubation of serum samples was performed according to EUROIMMUN immunoblotting instructions (eg in DPA-Dx Pollen 1, DP 3210-1601-1E). All incubation and washing steps were done at room temperature (+18°...

Embodiment 3

[0151] Example 3: A study based on a larger patient cohort using line blots and ELISA

[0152] patient selection

[0153] To assess the diagnostic value of different peanut storage proteins, adults with suspected peanut allergy based on history and / or sensitization (who had undergone diagnostic open or Double-blind placebo-controlled food challenge (DBPCFC) (n=127). Residual serum after routine blood collection one year before and after the food challenge date was available for 95 subjects. From this cohort, 40 peanut-allergic patients and 40 tolerant patients (using positive and negative challenges, respectively) were selected based on the availability of ImmunoCAP ISAC sensitization data. DBPCFC was performed following an international consensus protocol as previously described (Taylor SL, Hefle SL, Bindslev-Jensen C, Atkins FM, Andre C, Bruijnzeel-Koomen C et al. A consensus protocol for the determination of the threshold doses for allergic foods: How much is too much? C...

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Abstract

The invention relates to an improved assay for the diagnosis of peanut allergy. The present invention relates to a diagnostically useful carrier comprising a means for specifically capturing an antibody to Ara h 7 isotype 7.0201 in a sample from a subject; a method comprising the step detecting in a sample from a subject the presence or absence of an antibody to Ara h 7 isotype 7.0201 and a pharmaceutical composition comprising Ara h 7 isotype 7.0201 or a variant thereof.

Description

[0001] This application is a divisional application, the application number of its parent application is 201711468054.0, the application date is December 29, 2017, the earliest priority date is February 15, 2017, and the invention name is "Improved diagnosis of peanut allergy measurement method". technical field [0002] The present invention relates to a diagnostically useful carrier comprising means for specifically capturing antibodies against Ara h 7 isoform 7.0201 in a sample from a subject; including detection of antibodies against Ara h in a sample from a subject 7. Methods of steps of the presence or absence of antibodies of isoform 7.0201 and pharmaceutical compositions comprising Ara h 7 isoform 7.0201 or variants thereof. Background technique [0003] Among food allergies, those associated with peanuts (Arachis hypogea) have received the most research attention because they are relatively common, often permanent, and often severe. The Food Allergy Mortality Regis...

Claims

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Application Information

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IPC IPC(8): G01N33/68C07K14/415C07K16/16
CPCC07K14/415C07K16/16G01N33/6893G01N2333/415G01N2800/24G01N33/6854A61K39/35A61K2039/577A61P37/08C07K2319/00
Inventor W·苏尔S·罗威尔B·布瑞克斯A·韦曼
Owner EUROIMMUN MEDIZINISCHE LABORDIAGNOSTIKA
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