title: Assessment of chemokine profiles in human skin biopsies by an immunoaffinity capillary electrophoresis chip.
abstract: Atopic dermatitis is a skin condition resulting in a skin rash from exposure to environmental factors. Skin biopsies taken from patients suffering from atopic dermatitis were micro-dissected and analyzed using a microchip-based immunoaffinity CE system for the presence of CXCL1, CXCL5 and CXCL8 and CCL1, CCL3 and CCL5 chemokines. Disposable immunoaffinity disks with immobilized antibodies were used to capture the CXC and CC chemokines from the homogenized skin samples. The captured analytes were then labeled with AlexaFluor 633, eluted from the disk and separated by CE. The labeled chemokines were identified and quantified by laser induced fluorescence. The total analysis time was less than 40min, including the biopsy microdissection, pre-analysis preparation of the sample and the ICE-CHIP analysis, which took less than 10min with inter- and intra-assay CV's below 6.4%. Microchip-based immunoaffinity CE could distinguish between normal skin biopsies and those with inflammation. Patients with neutrophil cellular infiltrates by histopathology showed increased concentrations of CXCL1, CXCL5 and CXCL8 while increases of CCL1, CCL3 and CCL5 corresponded to the patient group demonstrating monocytic and T-lymphocyte infiltration by histopathology. This system demonstrates the ability to identify and quantify immunochemical analytes in frozen sections taken from clinical histopathology samples.
authors.affiliation: Micro Analytical Immunochemistry Unit, Biomedical Engineering and Physical Science Shared Resource, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Building 13, Room 3E41, 13 Center Drive, Bethesda, MD 20817, USA. KalishH@mail.nih.gov
authors.forename: Heather
authors.initials: H
authors.lastname: Kalish
authors.forename: Terry M
authors.initials: TM
authors.lastname: Phillips
mesh_terms: D000328:Adult; D055910:Antibodies, Immobilized / Q000737:chemistry; D001706:Biopsy; D016022:Case-Control Studies; D018925:Chemokines / Q000032:analysis* / Q000737:chemistry; D003876:Dermatitis, Atopic / Q000473:pathology; D019075:Electrophoresis, Capillary / Q000295:instrumentation* / Q000379:methods; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods; D007202:Indicators and Reagents / Q000737:chemistry; D008214:Lymphocytes / Q000737:chemistry; D040081:Protein Array Analysis / Q000295:instrumentation* / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012867:Skin / Q000737:chemistry* / Q000473:pathology; D013997:Time Factors
chemical_list: D055910:Antibodies, Immobilized; D018925:Chemokines; D007202:Indicators and Reagents