Analysis of secretory immunoglobulin A in human saliva by laser-induced fluorescence capillary electrophoresis.
The utility of capillary electrophoresis (CE) has been demonstrated for the analysis of secretory immunoglobulin A (sIgA) in human saliva. The amount of sIgA in saliva correlates with immune status. For detecting salivary sIgA, laser-induced fluorescence was conducted in this report for signal amplification. sIgA and anti-sIgA antibody were labeled with cyanine fluorescence (Cy5) for competitive immunoassay and non-competitive analysis, respectively. Cy5 was excited by He-Ne laser with a wavelength of 635 nm, with maximum emission at 670 nm. Migration time during electrophoresis depended on whether sIgA-Cy5 was mixed with antibody or anti-sIgA-Cy5 mixed with sIgA to form Ag-Ab complex. The results indicated that CE competitive immunoassay was effective for analyzing serum sIgA, but not for salivary sIgA. However, salivary sIgA can be analyzed by complex formation assay. The peak area of the complex was proportional to the amount of sIgA added. A standard linear regression curve was generated using purified sIgA. From this standard curve, the amount of sIgA from saliva of either normal or immunocompromised patients can be calculated from the Ag-Ab complex peak area.
Liu
Cheng-Ming
CM
Institute of Biomedical Technology, Taipei Medical University, 250 Wu-Shin Street, Taipei, Taiwan. cheng-m@tmu.edu.tw
Tung
Kuo-Hua
KH
Chang
Tsui-Hua
TH
Chien
Chen-Chin
CC
Yen
Mao-Hsiung
MH
D002021:Buffers; D019075:Electrophoresis, Capillary / Q000379:methods*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007071:Immunoglobulin A, Secretory / Q000032:analysis*; D007834:Lasers; D012463:Saliva / Q000737:chemistry*; D013050:Spectrometry, Fluorescence / Q000379:methods*
D002021:Buffers; D007071:Immunoglobulin A, Secretory