title: Capillary isoelectric focusing immunoassay as a new nanoscale approach for the detection of oligoclonal bands.
abstract: The detection of oligoclonal bands (OCBs) in cerebrospinal fluid is an indicator of intrathecal synthesis of immunoglobulins which is a neurochemical sign of chronic inflammatory brain diseases. Intrathecally synthesized IgGs are typically observed in patients with multiple sclerosis. The current standard protocol for the detection of OCBs is IEF on agarose or polyacrylamide gels followed by immunoblotting or silver staining. These methods are time consuming, show substantial interlaboratory variation and cannot be used in a high throughput-approach. We have developed a new nanoscale method for the detection of OCBs based on automated capillary IEF followed by immunological detection. Evidence for intrathecal IgG synthesis was found in all tested patients (n = 27) with multiple sclerosis, even in two subjects who did not have oligoclonal bands according to standard methods. The test specificity was at 97.5% (n = 19). Our findings indicate that the novel OCB-CIEF-immunoassay is suitable for the rapid and highly sensitive detection of OCBs in clinical samples. Furthermore, the method allows for a higher sample throughput than the current standard methods.
authors.affiliation: Department of Neurology, University of Ulm, Germany.
authors.forename: Steffen
authors.initials: S
authors.lastname: Halbgebauer
authors.forename: Ute
authors.initials: U
authors.lastname: Haußmann
authors.forename: Hans
authors.initials: H
authors.lastname: Klafki
authors.forename: Hayrettin
authors.initials: H
authors.lastname: Tumani
authors.forename: Jens
authors.initials: J
authors.lastname: Wiltfang
authors.forename: Markus
authors.initials: M
authors.lastname: Otto
mesh_terms: D000328:Adult; D016022:Case-Control Studies; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005260:Female; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods*; D015151:Immunoblotting / Q000379:methods; D007074:Immunoglobulin G / Q000096:biosynthesis; D007249:Inflammation / Q000134:cerebrospinal fluid / Q000276:immunology; D007525:Isoelectric Focusing / Q000379:methods*; D057230:Limit of Detection; D008297:Male; D008875:Middle Aged; D009103:Multiple Sclerosis / Q000134:cerebrospinal fluid / Q000276:immunology; D036103:Nanotechnology / Q000295:instrumentation / Q000379:methods; D043025:Oligoclonal Bands / Q000134:cerebrospinal fluid*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D007074:Immunoglobulin G; D043025:Oligoclonal Bands
keywords: Capillary IEF; Intrathecal IgG synthesis; Multiple sclerosis; Oligoclonal bands