Detection of human immunoglobulin in microchip and conventional capillary electrophoresis with contactless conductivity measurements.
The detection of human immunoglobulin M (IgM) was performed using capacitively coupled contactless conductivity detection (CCD) in electrophoresis carried out in conventional capillaries as well as on glass and poly(meth-yl methacrylate) (PMMA) microdevices. Also achieved was the analyses of IgG (an anti-human IgM) and the complex formed in the reaction between the two immunoreagents. It is demonstrated that CCD is a powerful tool suitable not only for the detection of antibodies but also for monitoring an immunological interaction. Conductivity measurements allow the direct determination of immunoreagents, and it is advantageous, since no labels are required. The immunoglobulin IgM has been taken as model analyte. The reproducibility of the analytical signal (RSD = 1%), sensitivity and limits of detection obtained for IgM (0.15 ng/mL in conventional capillaries and 34 ng/mL in microchips) are comparable to those previously obtained with amperometric detection. The immunological reaction was performed either in conventional microtiter plates as used in ELISA or in situ on the glass chip.
Departamento de Química Física y Analítica, Universidad de Oviedo, Asturias, Spain.
Eva M
EM
Abad-Villar
Jatisai
J
Tanyanyiwa
M Teresa
MT
Fernández-Abedul
Agustín
A
Costa-García
Peter C
PC
Hauser
D004553:Electric Conductivity; D019075:Electrophoresis, Capillary / Q000295:instrumentation* / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D005898:Glass; D006801:Humans; D007118:Immunoassay; D007074:Immunoglobulin G / Q000032:analysis*; D007075:Immunoglobulin M / Q000032:analysis*; D007526:Isoelectric Point; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors
D007074:Immunoglobulin G; D007075:Immunoglobulin M