ATTOMOLAR PROTEIN DETECTION IN COMPLEX DAMLE MATRICES WITH SEMI-HOMOGENEOUS FLUIDIC FORCE DISCRIMINATION ASSAYS

摘要

A semi-homogenous fluidic force discrimination assay method wherein micron- sized beads and any required intermediate receptors (e.g. secondary antibodies) are first mixed directly with a sample, allowing target analytes to be efficiently captured onto the beads. The target- loaded beads are then specifically captured onto a microarray surface, with nonspecifically bound beads removed by controlled, laminar fluidic forces. Also disclosed is a surface chemistry method for attaching NeutrAvidin to a nitride surface. A surface having primary amines the surface is reacted with succinimidyl 4-formylbenzoate (SFB). NeutrAvidin is reacted with succinimidyl 4-hydrazinonicotinate acetone hydrazone (SANH). The SFB-reacted surface is incubated with the NA-SANH solution under acidic conditions to configure said surface with stable SFB-SANH. The stable surface is stored at 4° C and remains stable for at least 4 weeks.