Microarray technology applied to human allergic disease

Research output: Contribution to journalReview article

Abstract

IgE antibodies serve as the gatekeeper for the release of mediators from sensitized (IgE positive) mast cells and basophils following a relevant allergen exposure which can lead to an immediate-type hypersensitivity (allergic) reaction. Purified recombinant and native allergens were combined in the 1990s with state of the art chip technology to establish the first microarray-based IgE antibody assay. Triplicate spots to over 100 allergenic molecules are immobilized on an amine-activated glass slide to form a single panel multi-allergosorbent assay. Human antibodies, typically of the IgE and IgG isotypes, specific for one or many allergens bind to their respective allergen(s) on the chip. Following removal of unbound serum proteins, bound IgE antibody is detected with a fluorophore-labeled anti-human IgE reagent. The fluorescent profile from the completed slide provides a sensitization profile of an allergic patient which can identify IgE antibodies that bind to structurally similar (cross-reactive) allergen families versus molecules that are unique to a single allergen specificity. Despite its ability to rapidly analyze many IgE antibody specificities in a single simple assay format, the chip-based microarray remains less analytically sensitive and quantitative than its singleplex assay counterpart (ImmunoCAP, Immulite). Microgram per mL quantities of allergen-specific IgG antibody can also complete with nanogram per mL quantities of specific IgE for limited allergen binding sites on the chip. Microarray assays, while not used in clinical immunology laboratories for routine patient IgE antibody testing, will remain an excellent research tool for defining sensitization profiles of populations in epidemiological studies.

Original languageEnglish (US)
Article number3
JournalMicroarrays
Volume6
Issue number1
DOIs
StatePublished - Mar 1 2017

Fingerprint

Allergens
Microarrays
Antibodies
Immunoglobulin E
Technology
Assays
Immunology
Immunoglobulin G
Molecules
Fluorophores
Immediate Hypersensitivity
Binding sites
Antibody Specificity
Basophils
Allergy and Immunology
Mast Cells
Amines
Glass
Blood Proteins
Epidemiologic Studies

Keywords

  • Allergen extract
  • Component resolved diagnosis
  • Human
  • IgE
  • Immunoenzymetric assay
  • Immunosorbent allergen chip
  • ISAC
  • Microarray
  • Molecular allergen
  • Serodiagnosis

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Bioengineering
  • Biomedical Engineering

Cite this

Microarray technology applied to human allergic disease. / Hamilton, Robert G.

In: Microarrays, Vol. 6, No. 1, 3, 01.03.2017.

Research output: Contribution to journalReview article

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