Allergen arrays for antibody screening and immune cell activation profiling generated by parallel lipid dip-pen nanolithography

摘要

Multiple-allergen testing for high throughput and high sensitivity requires the development of miniaturized immunoassays that allow for a large test area and require only a small volume of the test analyte, which is often available only in limited amounts. Developing such miniaturized biochips containing arrays of test allergens needs application of a technique able to deposit molecules at high resolution and speed while preserving its functionality. Lipid dip-pen nanolithography (L-DPN) is an ideal technique to create such biologically active surfaces, and it has already been successfully applied for the direct, nanoscale deposition of functional proteins, as well as for the fabrication of biochemical templates for selective adsorption. The work presented here shows the application of L-DPN for the generation of arrays of the ligand 2,4-dinitrophenyl[1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-[6-[(2, 4-dinitrophenyl)amino]hexanoyl] (DNP)] onto glass surfaces as a model system for detection of allergen-specific Immunoglobin E (IgE) antibodies and for mast cell activation profiling. Multiple-allergen testing at high throughput and high sensitivity requires the development of miniaturized immunoassays that would allow for large test areas and require only a small volume of the test analyte. Application of lipid dip-pen nanolithography for the generation of arrays of the ligand 2,4-dinitrophenyl[1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-[6- [(2,4-dinitrophenyl)amino]hexanoyl] (DNP)] onto glass surfaces is presented as a model system for the detection of allergen-specific IgE (immunoglobulin E) antibodies and for mast cell activation profiling.