title: Surface immobilized hydrogels as versatile reagent reservoirs for microarrays.
abstract: The accuracy of antibody-based microarrays depends on eliminating or at least minimizing the effect of cross-reactive components. In this work, a method to create a versatile, compartmentalized storage system for antibodies and soluble microarray reagents is described. These containers are made of hydrogel plugs immobilized on a slide facing the corresponding sub-array on a printed surface. The hydrogels were polymerized using a 4% w/v N,N-dimethylacrylamide and 3% w/v bisacrylamide solution, thus generating large pore sizes to allow the facile transfer of intact, functional biomolecules. The hydrogel microenvironments can be desiccated and rehydrated with a desired solution, and they can store reagents in a dry form. These hydrogels were shown to provide limits of detection similar to those obtained by conventional incubation conditions in an assay for HIV p24 antigen. Moreover, they prevent cross-reactivity issues in the detection of Staphylococcus aureus enterotoxins A and B and promote the accurate quantification of interleukin 10 by a microarray-integrated calibration curve.
authors: Consiglio Nazionale delle Ricerche, Istituto di Chimica del Riconoscimento Molecolare (ICRM CNR), Milano, Italy.
authors: Laura
authors: L
authors: Sola
authors: Paola
authors: P
authors: Gagni
authors: Marina
authors: M
authors: Cretich
authors: Marcella
authors: M
authors: Chiari
mesh_terms: D000178:Acrylamides / Q000737:chemistry*; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D002138:Calibration; D003429:Cross Reactions; D003890:Desiccation; D004768:Enterotoxins / Q000032:analysis / Q000276:immunology; D004867:Equipment Design; D016655:HIV Core Protein p24 / Q000032:analysis / Q000276:immunology; D020100:Hydrogels; D007118:Immunoassay / Q000295:instrumentation* / Q000592:standards; D016753:Interleukin-10 / Q000032:analysis / Q000276:immunology; D057230:Limit of Detection; D058105:Polymerization; D016062:Porosity; D040081:Protein Array Analysis / Q000295:instrumentation* / Q000592:standards; D015203:Reproducibility of Results
chemical_list: D000178:Acrylamides; D000906:Antibodies; D004768:Enterotoxins; D016655:HIV Core Protein p24; D020100:Hydrogels; D016753:Interleukin-10; C026066:enterotoxin A, Staphylococcal; C031818:enterotoxin B, staphylococcal; C099046:N,N-dimethylacrylamide