Method for allergen characterization
OwnerThomas A. Beardslee, Michael G. Zeece +2inventors
InventorsThomas A. Beardslee, Michael G. Zeece, Guatam Sarath, John P. Markwell
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Abstract
Disclosed are methods for the characterization of allergens using recombinant fusion protein in an enzyme-linked immunosorbent (ELISA) assay. Also disclosed are methods for determining allergen sensitivity and methods for immunotherapy using the ELISA of the present invention.
Claims26
A method for allergen characterization comprising:
- a) obtaining a recombinant fusion protein expressed by a host cell, said recombinant fusion protein containing a first amino acid sequence of a known or suspected allergen or allergen fragment fused to a second amino acid sequence native to the host cell expressing said recombinant fusion protein;
- b) attaching said recombinant fusion protein to a substrate through said native protein;
- c) contacting said recombinant fusion protein attached to said substrate with a biological sample from an individual;
- and
- d) detecting the binding of immunoglobulin E molecules in said biological sample to said recombinant fusion protein.
A method for allergen characterization comprising:
- a) obtaining a recombinant fusion protein expressed by an E coli bacterium, said recombinant fusion protein containing a first amino acid sequence of a known or suspected allergen or allergen fragment fused to a second amino acid sequence of thioredoxin;
- b) attaching said recombinant fusion protein to a substrate by binding said thioredoxin with an antibody attached to said substrate;
- c) contacting said recombinant fusion protein attached to said substrate with a biological sample from an individual;
- and
- d) detecting the binding of immunoglobulin E molecules in said biological sample to said recombinant fusion protein.
A kit comprising a recombinant fusion protein obtained from a host cell, said recombinant fusion protein containing a first amino acid sequence of a known or suspected allergen or allergen fragment fused to a second amino acid sequence native to said host cell said recombinant fusion protein bound to a solid substrate through said native amino acid sequence;
- and instructions for using said recombinant fusion protein to determine IgE binding to said known or suspected allergen.
A method for epitope determinationcomprising:
- a) obtaining a recombinant fusion protein expressed by a host cell, said recombinant fusion protein containing a first amino acid sequence comprising a known or suspected epitope fused to a second amino acid sequence native to the host cell expressing said recombinant fusion protein;
- b) attaching said recombinant fusion protein to a substrate through said native protein;
- c) contacting said recombinant fusion protein attached to said substrate with a biological sample containing an immunoglobulin;
- and
- d) detecting the binding of immunoglobulin molecules in said biological sample to said recombinant fusion protein.
Dependencies are inferred from the claim text.