Allergen diagnosis microarray with high-density immobilization capacity using diamond-like carbon-coated chips for profiling allergen-specific IgE and other immunoglobulins.
The diagnosis of antibody-mediated allergic disorders is based on clinical findings, skin prick tests and detection of allergen-specific IgE in serum. Here, we present a new microarray technique of high-density antigen immobilization using carboxylated arms on the surface of a diamond-like carbon (DLC)-coated chip. High immobilization capacity of antigen on DLC chip at (0.94-7.82)×10(9) molecules mm(-2) allowed the analysis of allergen-specific immunoglobulins against not only purified proteins but also natural allergen extracts with wide assay dynamic range. The higher sensitivity of the allergen-specific IgE detection on DLC chip was observed for comparison with the UniCAP system: the DLC chip allowed lowering the limit of dilution rate in UniCAP system to further dilution at 4-8-fold. High correlations (ρ>0.9-0.85) of allergen-specific IgE values determined by the DLC chip and UniCAP were found in most of 20 different allergens tested. The DLC chip was useful to determine allergen-induced antibodies of IgA, IgG, IgG1, and IgG4 in sera, apart from IgE, as well as secretory IgA in saliva against the same series of allergens on the chip in a minimal amount (1-2 μL) of sample.
Division of Enzyme Chemistry, Institute for Enzyme Research, The University of Tokushima, Tokushima 770-8503, Japan.
Koichi
K
Suzuki
Mineyoshi
M
Hiyoshi
Hitomi
H
Tada
Miwa
M
Bando
Takao
T
Ichioka
Norio
N
Kamemura
Hiroshi
H
Kido
D000293:Adolescent; D000328:Adult; D000485:Allergens / Q000737:chemistry* / Q000276:immunology*; D000918:Antibody Specificity; D002648:Child; D002675:Child, Preschool; D018130:Diamond / Q000737:chemistry*; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007073:Immunoglobulin E / Q000032:analysis* / Q000097:blood / Q000276:immunology; D007223:Infant; D007231:Infant, Newborn; D046228:Microarray Analysis / Q000379:methods*; D015203:Reproducibility of Results; D012463:Saliva / Q000737:chemistry; D055815:Young Adult
D000485:Allergens; D007073:Immunoglobulin E; D018130:Diamond