title: Measurement of Inflammatory Chemokines in Micro-dissected Tissue Biopsy Samples by Chip-Based Immunoaffinity Capillary Electrophoresis.
abstract: 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. Electrophoretic separation of all of the analytes was achieved in 2.5 min with quantification of each peak being performed by online LIF detection and integration of each peak area. The degree of accuracy and precision achieved by the chip-based system is comparable to conventional immunoassays and the system is robust enough to be applied to the analysis of clinical samples. Further, with the expanding array of antibodies that are commercially available, this chip-based system can be applied to a wide variety of different biomedical and clinical analyses.
authors.affiliation: Microanalytical Immunochemistry Unit, Biomedical Engineering and Physical Science Shared Resource, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, 20814, USA. tmphil46@yahoo.com.|National Institute of Biomedical Imaging and Bioengineering, 4101 Albemarle Street NW Unit 508, Washington, DC, 20016, USA. tmphil46@yahoo.com.
authors.forename: Terry M
authors.initials: TM
authors.lastname: Phillips
authors.affiliation: Retired from National Institutes of Health, Bethesda, MD, USA.
authors.forename: Edward
authors.initials: E
authors.lastname: Wellner
authors.affiliation: Microanalytical Immunochemistry Unit, Biomedical Engineering and Physical Science Shared Resource, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, 20814, USA.
authors.forename: Shane
authors.initials: S
authors.lastname: McMohan
authors.affiliation: Microanalytical Immunochemistry Unit, Biomedical Engineering and Physical Science Shared Resource, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, 20814, USA.
authors.forename: Heather
authors.initials: H
authors.lastname: Kalish
mesh_terms: D001706:Biopsy; D016207:Cytokines / Q000302:isolation & purification*; D019075:Electrophoresis, Capillary / Q000379:methods*; D046209:Electrophoresis, Microchip / Q000379:methods*; D006801:Humans; D056656:Lab-On-A-Chip Devices; D042282:Microdissection; D012867:Skin / Q000276:immunology / Q000473:pathology*
chemical_list: D016207:Cytokines
keywords: Capillary electrophoresis; Chemokines; Human biopsies; Immunoaffinity; Micro-dissection; Microchip