Analysis of secretory IgA in human saliva by laser induced fluorescence capillary electrophoresis

• Main Authors: Tsui-Hua Chang, 張翠華
• Other Authors: Cheng-Ming Liu, Ph.D.
• Format: Others
• Language: zh-TW
• Published: 2001
• Online Access: http://ndltd.ncl.edu.tw/handle/38294861484500552218

碩士 === 台北醫學院 === 生物醫學技術研究所 === 89 === Abstract Saliva is extremely sensitive to changes in oral and systemic diseases. As a new role for clinical tool, saliva offers many advantages over serum, saliva is easy to collect, painless and non invasive. It can be used as an indicator on the status of individual immunity. Secretory immunoglobulin A (sIgA) is the most abundant immunoglobulin in saliva and plays an important role in mucosal immunity. sIgA measurement in saliva as an index of mucosal immunity, has repeatedly been shown sensitive to physiological variables. For instance increase level comes after bacterial or viral infections, contrary reduction level can be observed while under stressful circumstances. Capillary electrophoresis (CE) is a new analytical technique for assessment of different variety of body fluids, as it requ.ires only a small volume and provides a rapid analysis with high efficient separation. In addition the instrument is easy to be automated. This study aims to establish an optimal condition for CE analyzed trace amount of sIgA in saliva. Laser induced fluorescence (LIF) conducted in capillary electrophoresis improvement of its sensitivity, both of the competitive (CI-CE) and the non competitive (direct) immunoassays (NCI-CE). The fluorescence Cy5 on the maximum absorption at 649 nm and the maximum emission efficiency at 670 nm was used for either sIgA or anti-sIgA labeling. The electrophoresis was conducted under 15kV in borate buffer 150 mM with CHAPS, pH8.5. The method is based on the different mobility of the free and bound tracer (fluorescently labeled compound). The amounts of sIgA in graduate student''s saliva were assessed during the period under stress or stress free. The results indicate that the NCI-CE is more stable and reproducible than the CI-CE for sIgA quantitative analysis.