Hydrogel-Immobilized Antibodies for Microfluidic Immunoassays: <I>Hydrogel Immunoassay</I>

Abstract

The integration of immunoassays in microfluidic devices is a rapidly developing research area combining the power of immunoassays with the inherent benefits of microfluidics. Here, a general overview of microfluidic-based immunoassays is presented along with a method for immobilizing antibodies in polyacrylamide gel plugs set in microfluidic channels. These antigen-specific hydrogels can be rapidly formed by photopolymerizing monomer solutions mixed with antibodies or other large proteins. The resulting antigen-specific hydrogels contain pore sizes appropriate for physical entrapment of large antibodies while remaining permeable to smaller proteins. The open structure of these hydrogels enables the capture and concentration of target antigens present at low concentrations. Such physical entrapment provides a conceptually simple method of immobilization compared with immobilization of proteins on surfaces and offers advantages such as resistance to chemical and thermal denaturation.

Bioinformatic Approaches to the Identification of Novel Neuropeptide Precursors

“…Porous membranes provide a convenient support material, which is strong and for which a variety of materials and protocols are available. Hydrogels cannot compete with membranes in terms of strength and durability, but they provide the best 3D support for the immobilisation of test molecules (whether proteins or peptides) in their native functional state in highly porous hydrogel substrate suitable for both functional assays (36) and immunoassays (26,37,38). When hydrated, the hydrogels swell, allowing easy access for the molecules and short diffusion times, but when dried, the gel thickness is reduced significantly, resulting in focussing of the trapped fluorescence in a thinner layer.…”

“…Do not remove the probe from the beaker far in advance of surgery to prevent drying out of the membrane (it will become unusable). 38. The probe is still attached to the syringe through the inlet tubing, so the pump should be close enough that it can be connected to the crab without straining the tubing.…”

“…Further increases are counterproductive as the procedure becomes very long and sample evaporation becomes an issue. 38. High humidity should be maintained inside the robot whilst spotting, especially for longer runs.…”

“…. 15,18,21,[37][38]40,45,51,[53][54]67,77,80,88,[103][104][105]111,125,130,139,142,[148][149]197,[209][210][211]214,243,[249][250]253,256,280,171,174,181 Threshold . .…”

Identification and Relative Quantification of Neuropeptides from the Endocrine Tissues

“…38, 42, 80, 103, 108, 146, 148, 195, 199, 202 . 37-38, 40, 42-44, 83, 102, 193, 250 ESI Q-TOF [37][38]40,[43][44]83,102,193 EST .121, 131, 139-140, 192, 196 . 38, 119, 131, 139, 199-200, 261, .…”