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| Flowable demineralized bone powder composition and its use in bone repair |
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| Crosslinkable polypeptide compositions |
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Peanut allergens and methods
| Details |
Inventors: Burks, Jr., A. Wesley; Stanley, J. Steven; Bannon, Gary A.; Cockrell, Gael; Helm, Ricki M.;
Assignee: University of Arkansas (Little Rock, AR)
Primary Examiner: Scheiner; Laurie
Assistant Examiner:
Attorney, Agent or Firm: Choate, Hall & Stewart, Jarrell; Brenda H., Lyon; Charles E.
One of the major peanut allergens, Ara h I, was selected from cDNA expression library clones using Ara h I specific oligo-nucleotides and polymerase chain reaction technology. The Ara h I clone identified a 2.3 kb mRNA species on a Northern blot containing peanut poly A+RNA. DNA sequence analysis of the cloned inserts revealed that the Ara h I allergen has significant homology with the vicilin seed storage protein family found in most higher plants. The isolation of the Ara h I clones allowed the synthesis of this protein in E. coli cells and subsequent recognition of this. recombinant protein in immunoblot analysis using serum IgE from patients with peanut hypersensitivity. |
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DETAILED DESCRIPTION OF THE INVENTION In accordance with a preferred embodiment of the present invention, a monoclonal antibody (MAb) based enzyme-linked immunosorbent assay (ELISA) and method is provided for detecting and quantifying a major peanut allergen, Ara h I, in food products.
The Ara h I peanut allergen (63.
5 kD fraction) was used as an immunogen to produce monoclonal antibodies (MAbs) having specificity for the selected peanut allergen.
The Ara h I allergen was isolated as described in the Burks et al.
article entitled "Identification of a Major Peanut Allergen, Ara h I, In Patients With Atopic Dermatitis and Positive Peanut Challenges", Journal of Allergy Clinical Immunology 1991; 88:172-9.
Patients with atopic dermatitis (AD) who had positive double-blind, placebo controlled, food challenge to peanut were used in the process of identification and purification of the peanut allergens.
A major peanut allergen, Ara h I, was identified by use of anion-exchange column chromatography, sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), ELISA, thin layer isoelectric focusing (TLIEF) and IgE-specific immunoblotting.
The Ara h I allergen is a glycoprotein from a pure strain of Florunner peanuts that has a mean molecular weight of 63.
5 kD and a mean isoelectric point (pI) of 4.
55.
Nine patients (mean age 4.
2 years) with AD and a positive immediate prick skin test to peanut had either a positive DBPCFC or a convincing history of peanut anaphylaxis.
(The allergic reaction was potentially life-threatening, that is, laryngeal edema, severe wheezing and/or hypotension.
) Five milliliters of venous blood was obtained from each patient and allowed to clot, and then the serum was collected.
An equal volume of serum from each donor was mixed to prepare a nine-person, peanut-specific, IgE antibody (Ab) pool.
Three commercial lots of southeastern runners (Arachis hypogaea) (Florunner), medium grade peanuts from the 1979 crop (North Carolina State University), were used in this study
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