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. Electrophoretic separation of all of the analytes was achieved in 2.2 min with quantification of each peak being performed by online LIF detection and integration of each peak area. Comparison of the results obtained from the chip-based system to those obtained using commercially available high-sensitivity immunoassays demonstrated that the chip-based assay provides a fast, accurate procedure for studying the concentrations of inflammatory biomarkers in complex biological materials. The degree of accuracy and precision achieved by the chip-based CE is comparable to conventional immunoassays and the system is capable of analyzing circa six samples per hour. With the ever-expanding array of antibodies that are commercially available, this chip-based system can be applied to a wide variety of different biomedical analyses.
Phillips
Terry M
TM
Nanoscale Immunodiagnostics, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD 20892, USA. phillipt@mail.nih.gov
Wellner
Edward F
EF
D000906:Antibodies / Q000276:immunology; D015415:Biomarkers / Q000032:analysis; D001706:Biopsy; D019075:Electrophoresis, Capillary / Q000379:methods*; D046209:Electrophoresis, Microchip; D006801:Humans; D007118:Immunoassay; D007249:Inflammation / Q000276:immunology*; D012867:Skin / Q000378:metabolism*
D000906:Antibodies; D015415:Biomarkers