Performance of a polymer coated silicon microarray for simultaneous detection of food allergen-specific IgE and IgG4.
BACKGROUND
Microarray-based component-resolved diagnostics (CRD) has become an accepted tool to detect allergen-specific IgE sensitization towards hundreds of allergens in parallel from one drop of serum. Nevertheless, specificity and sensitivity as well as a simultaneous detection of allergen-specific IgG4 , as a potential parameter for tolerance development, remain to be optimized.


OBJECTIVE
We applied the recently introduced silicon chip coated with a functional polymer named copoly(DMA-NAS-MAPS) to the simultaneous detection of food allergen-specific IgE and IgG4 , and compared it with ImmunoCAP and ImmunoCAP ISAC. Inter- and intraslide variation, linearity of signal and working range, sensitivity and application of internal calibrations for IgE and IgG4 were assessed.


METHODS
Native and recombinant allergenic proteins from hen's egg and cow's milk were spotted on silicon chips coated with copoly(DMA-NAS-MAPS) along with known concentrations for human IgE and IgG4 . A serum pool and 105 patient samples were assessed quantitatively and semi-quantitatively with the ImmunoCAP and ImmunoCAP ISAC and correlated with IgE- and IgG4 -specific fluorescence on silicon microarrays.


RESULTS
Allergen-specific IgE and IgG4 were detected in parallel using two fluorescent dyes with no crosstalk. Results from the ImmunoCAP correlated better with microarray fluorescence than with ImmunoCAP ISAC except for the allergen ovomucoid. The working range of the silicon microarray for total hen's egg-specific IgE was comparable to the range of 0.1 to >100 kUA /L of the ImmunoCAP system, whereas for total cow's milk, the silicon microarray was less sensitive. Detectable allergen-specific IgG4 could be determined only for low concentrations, but still correlated positively with ImmunoCAP results.


CONCLUSIONS
We confirmed the ability of the polymer coated silicon microarray to be comparably sensitive to the ImmunoCAP ISAC for various food allergens. This suggests that the copoly(DMA-NAS-MAPS) microarray is a low-cost, self-producible alternative to the commercial ImmunoCAP ISAC in allergy research.
['Sievers|S|S|http://orcid.org/0000-0002-4265-1974', 'Cretich|M|M|', 'Gagni|P|P|', 'Ahrens|B|B|', 'Grishina|G|G|', 'Sampson|H A|HA|', 'Niggemann|B|B|', 'Chiari|M|M|', 'Beyer|K|K|']
D021181:Egg Hypersensitivity D005260:Female D006801:Humans D007073:Immunoglobulin E D007074:Immunoglobulin G D008297:Male D016269:Milk Hypersensitivity D040081:Protein Array Analysis D012825:Silicon
IgE IgG4 component-resolved diagnostics Immunologic tests allergens and epitopes clinical immunology food allergy protein microarray