Analysis of Inflammatory Mediators in Newborn Dried Blood Spot Samples by Chip-Based Immunoaffinity Capillary Electrophoresis.

A chip-based immunoaffinity capillary electrophoresis (ICE) system has been developed for measuring inflammatory mediators in dried blood samples routinely taken from newborn babies. A defined area of each dried blood spot was removed from the sample card and its contents eluted. The recovered eluates were injected into the chip and the analytes of interest isolated by the immunoaffinity disk within the chip.

Measurement of Inflammatory Chemokines in Micro-dissected Tissue Biopsy Samples by Chip-Based Immunoaffinity Capillary Electrophoresis.

To aid in the biochemical analysis of human skin biopsies, a chip-based immunoaffinity capillary electrophoresis (ICE) system has been developed for measuring inflammatory chemokines in micro-dissected areas of the biopsy. Following isolation of the areas of interest, the tissue was solubilized and the analytes of interest were isolated by the immunoaffinity disk within the chip. The captured analytes were labeled in situ with a 635 nm light-emitting laser dye and electro-eluted into the chip separation channel.

Detection of cerebral spinal fluid-associated chemokines in birth traumatized premature babies by chip-based immunoaffinity CE.

A major concern in treating premature infants with birth-associated head trauma is the rapid determination of reliable biomarkers of neuroinflammation. To this end a chip-based immunoaffinity CE device has been applied to determine the concentrations of inflammation-associated chemokines in samples of cerebral spinal fluid collected from such subjects. The chip utilizes replaceable immunoaffinity disks, to which reactive antibody fragments (FAb) of six antichemokine-specific antibodies were immobilized.

Receptor affinity CE for measuring bioactive inflammatory cytokines in human skin biopsies.

A chip-based receptor affinity CE system has been employed to measure the concentrations of bioactive pro-inflammatory cytokines in biopsy materials obtained from human atopic skin lesions. The device employs a replaceable affinity disk to which recombinant cytokine receptors have been chemically immobilized. Homogenates obtained from micro-dissected human skin samples were injected into the system where the bioactive cytokines were captured in the receptor affinity port and labeled in situ with a laser dye.

Chip-based immunoaffinity CE: application to the measurement of brain-derived neurotrophic factor in skin biopsies.

A chip-based immunoaffinity CE system has been employed to measure the concentrations of brain-derived neurotrophic factor in human skin biopsies, taken during atopic inflammatory events. The device employs a replaceable immunoaffinity disk to which capture antibodies have been chemically immobilized. Homogenates obtained from micro-dissected human skin samples were injected into the system, where the analyte of interest was captured in the immunoextraction port, thus allowing non-reactive materials to be removed prior to analysis.

A chip-based immunoaffinity capillary electrophoresis assay for assessing hormones in human biological fluids.

A chip-based capillary electrophoresis system has been designed for assessing the concentrations of four hormones in whole human blood, saliva, and urine. The desired analytes were isolated by immunoextraction using a panel of four analyte-specific antibodies immobilized onto a glass fiber insert within the injection port of the chip. Following extraction, the captured analytes were labeled prior to electro-elution into the chip separation channel, where they were resolved into four individual peaks in circa 2 min.

Analysis of inflammatory biomarkers from tissue biopsies by chip-based immunoaffinity CE.

To aid in the biochemical analysis of human skin biopsies, a semiautomatic chip-based CE system has been developed for measuring inflammatory biomarkers in microdissected areas of the biopsy. Following solubilization of the dissected tissue, the desired biomarkers were isolated by immunoaffinity capture using a panel of 12 antibodies, immobilized on a disposable glass fiber disk, within the extraction port of the chip. The captured analytes were labeled with a 635 nm light-emitting laser dye and electroeluted into the separation channel.

Measurement of naproxen in human plasma by chip-based immunoaffinity capillary electrophoresis.

An electrokinetic immunoassay performed in a chip-based capillary electrophoresis system is described for the rapid measurement of naproxen in human plasma. The system employs a fluorescently labeled antibody to capture and detect the analyte of interest within a 5 min total assay time with an LOD of 0.025 microg/mL and a saturation level of 450 microg/mL.

Measurement of neuropeptides in clinical samples using chip-based immunoaffinity capillary electrophoresis.

The current interest in micro-fabrication has extended to the clinical arena where there is a growing lobby for promoting these for point-of-care purposes. The advantages of such devices are their relative speed of analysis, lower reagent costs, and their application to clinical screening and diagnosis. Two chip-based capillary electrophoresis systems have been designed and their performance evaluated for rapidly measuring the concentrations of inflammatory neuropeptides in tissue fluids of patients with neuropeptide-associated muscle pain.

Development of a two-color laser fluorescence detector. On-line detection of internal standards and unknowns by capillary electrophoresis within the same sample.

A two-laser, two-color detector has been developed for the simultaneous detection of naturally occurring and recombinant (internal standards) cytokines within the same biological sample. The internal standards were labeled with Bimane and detected with a 408 nm laser while the natural cytokines were labeled with AlexaFluor633 and detected with a 633 nm laser. The two resulting electropherograms were plotted as overlaid traces and quantification of the natural materials determined by comparison with the standards.