Allergen diagnosis microarray with high-density immobilization capacity using diamond-like carbon-coated chips for profiling allergen-specific IgE and other immunoglobulins

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• 作者：Koichi  ; Suzuki^a ; Mineyoshi  ; Hiyoshi^a ; Hitomi  ; Tada^a ; Miwa  ; Bando^a ; Takao  ; Ichioka^b ; Norio  ; Kamemura^a ; Hiroshi  ; Kido^a ;   ; ^{kido@ier.tokushima-u.ac.jp}
• 关键词：Allergen ; Diamond-like carbon ; Immunoassay ; Microarray technology
• 刊名：Analytica Chimica Acta
• 出版年：2011
• 出版时间：14 November 2011
• 年：2011
• 卷：706
• 期：2
• 页码：321-327
• 全文大小：1210 K

文摘

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?.82) ¡Á 10⁹ molecules mm^? 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?-fold. High correlations (¦Ñ > 0.9?.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? ¦ÌL) of sample.