Dm. Paquette et Pr. Banks, Detection of specific antibodies using immunosubtraction and capillary electrophoresis instrumentation, ELECTROPHOR, 22(12), 2001, pp. 2391-2397
Solution-phase immunoassays based on capillary electrophoresis (CE) separat
ions have been shown to be rapid and simple to perform. The potential for s
ample matrix interference and incompatibility with multiplexing conditions
for antibody detection when dealing with real samples, however, has prompte
d the development of an assay that utilizes an immunosubtraction methodolog
y. A model assay for the detection of specific antibodies that relies on so
lid-phase extraction, CE and laser-induced fluorescence (LIF) detection is
described. The method, called immunocapture-immunosubtraction (ICIS), incor
porates an antibody capture/purification protocol using magnetic particles.
The detection of specific antibodies is achieved by CE-LIF analysis of a p
robe solution following incubation with the captured antibodies. As an exam
ple of the ICIS assay's capabilities, the relative quantification of anti-f
luorescein in serum is presented.