Rapid analysis of inflammatory cytokines in cerebrospinal fluid using chip-based immunoaffinity electrophoresis.
A chip-based capillary electrophoresis system has been designed for rapidly measuring the concentrations of inflammatory cytokines in the cerebrospinal fluid of patients with head trauma. Isolation of the reactive cytokines was achieved by immunoaffinity capture using a panel of six immobilized antibodies, directly attached to the injection port of the chip. The captured cytokines were labeled in situ with a red light-emitting laser dye and electroeluted into the separation channel. Separation of the isolated cytokines was achieved by electrophoresis in under 2 min with quantification of the resolved peaks being achieved by on-line laser-induced fluorescence and integration of each peak area. Comparison of the results to commercially available high-sensitivity immunoassays demonstrates that the chip-based assay provides a fast, accurate procedure for studying the concentrations of these analytes in complex biological materials. The degree of accuracy and precision achieved by the chip-based CE is comparable to conventional immunoassays, the system being able to analyze between 10-12 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 analyses.
Phillips
Terry M
TM
Ultramicro Analytical Immunochemistry Resource, Division of Bioengineering & Physical Science, Office of Research Services, National Institutes of Health, 13/3E42 9000 Rockville Pike, Bethesda, MD 20892, USA. phillipt@mail.nih.gov
D000906:Antibodies / Q000737:chemistry; D006259:Craniocerebral Trauma / Q000378:metabolism*; D016207:Cytokines / Q000032:analysis* / Q000134:cerebrospinal fluid; D019075:Electrophoresis, Capillary / Q000295:instrumentation*; D006801:Humans; D013050:Spectrometry, Fluorescence / Q000379:methods*
D000906:Antibodies; D016207:Cytokines