Detection of allergen specific immunoglobulins by microarrays coupled to microfluidics.
Allergen microarrays are under development for a component-resolved diagnosis of Type I (IgE-mediated) allergies. Here we report an improved microarray coupled to microfluidics for the detection of allergen specific immunoglobulin E (IgE). The signal intensity for IgE detection in serum has been improved by using glass slides coated with a novel poly[DMA-co-NAS] brush copolymer which is able to immobilize allergens in their native conformation and by carrying out the incubation step in dynamic conditions. The assay, fully automated, was performed in a microcell, using a software-controlled fluidic processor, to bring assay reagents on the surface of the array. Microfluidics turns the binding between serum immunoglobulins and immobilized allergens from a diffusion-limited to a kinetic-limited process by ensuring an efficient mixing of serum samples on the surface of the microarray. As a result of this, the binding of high affinity IgE antibodies is enhanced whereas that of low affinity IgG antibodies, which are present at higher concentration, is impaired paving the way to more accurate and sensitive results.
Cretich
Marina
M
Istituto di Chimica del Riconoscimento Molecolare (ICRM), CNR, Milano, Italy. marina.cretich@icrm.cnr.it
Di Carlo
Gabriele
G
Giudici
Cinzia
C
Pokoj
Sven
S
Lauer
Iris
I
Scheurer
Stephan
S
Chiari
Marcella
M