Detection of specific antibodies using immunosubtraction and capillary electrophoresis instrumentation.
Solution-phase immunoassays based on capillary electrophoresis (CE) separations have been shown to be rapid and simple to perform. The potential for sample matrix interference and incompatibility with multiplexing conditions for antibody detection when dealing with real samples, however, has prompted the development of an assay that utilizes an immunosubtraction methodology. A model assay for the detection of specific antibodies that relies on solid-phase extraction, CE and laser-induced fluorescence (LIF) detection is described. The method, called immunocapture-immunosubtraction (ICIS), incorporates an antibody capture/purification protocol using magnetic particles. The detection of specific antibodies is achieved by CE-LIF analysis of a probe solution following incubation with the captured antibodies. As an example of the ICIS assay's capabilities, the relative quantification of anti-fluorescein in serum is presented.
Paquette
D M
DM
Department of Chemistry and Biochemistry, Concordia University, Montreal, Quebec, Canada. dm_paqu@alcor.concordia.ca
Banks
P R
PR
D000818:Animals; D000906:Antibodies / Q000032:analysis*; D000918:Antibody Specificity; D000936:Antigen-Antibody Complex / Q000032:analysis; D002138:Calibration; D019075:Electrophoresis, Capillary / Q000295:instrumentation* / Q000379:methods; D005240:Feasibility Studies; D019793:Fluorescein / Q000032:analysis; D005456:Fluorescent Dyes / Q000032:analysis; D005470:Fluorometry; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods*; D018189:Immunomagnetic Separation; D007163:Immunosorbent Techniques; D007834:Lasers; D011817:Rabbits; D013382:Subtraction Technique
D000906:Antibodies; D000936:Antigen-Antibody Complex; D005456:Fluorescent Dyes; D019793:Fluorescein