Construction And Clinical Investigation Of Monoclonal Antidody Reagent Of Anti-human Secretory Component Of SIgA

Objective First, we obtained secretory component(SC) by purifying colostrums; Then, we produced anti-SC PAb by Utilizing SC to immune rabbit; Third, we resuscitated and distensibly cultivated hybridoma cell strains of anti-human SC mAb to produce this antibody. Finally, we utilized the high specificity of mAb and high avidity of PAb to construct mAb ELISA reagent. We use this reagent to study SIgA of Cervical secretions of cervical cancer patients. Moreover, we have further studied SIgA level of viral Hepatitis.Methods Our investigation have three parts: ( 1 ) separation, purification andidentification of SIgA and SC; (2) Construction of anti-SC mAb ELISA reagent; (3 ) The clinical investigations of anti-human SC mAb ELISA reagent.Results We can obtain SIgA and SC from colostrums by gel filtration, ion exchange chromatography and salt fraction. Through Double immunodiffusion, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western Blot, we can more confirm the products are SIgA and SC. The molecular weight of SC is 75 kD. We produced anti-SC PAb by Utilizing purified SC to immune rabbit, it was peculiarly reacting with SC antigen. We produced anti-human SC mAb4G3 .Its valence (1:4000) can be detected by ELISA . We utilized the high specificity of mAb and high avidityof PAb to construct mAb ELISA reagent. The best configuration of this reagent was: Anti-human SC PAb was considered to be solid phase antibody; SIgA was detected; anti-human SC mAb4G3 was considered to be Enzyme labeled antibody. We can evaluated this reagent in sensitivity, specificity, Stabilizing curve slope. Its Clinical investigations indicate that: fistly, SIgA level of Cervical secretions of normal woman is higher than that of woman who suffered from cervical cancer; Secondary, the lower the level of SIgA, the more serious the suffer degree of viral hepatitis patients. But all of them are significantly higher than that of normal persons.Conclusions: This investigation indicates that, if we considered Anti-human SC PAb as solid phase antibody; then labeled anti-human SC mAb, we can construct anti-SC mAb ELISA reagent which has high specificity and high avidity. Furthermore, this anti-human SC mAb ELISA reagent can be apply in the investigation of several clinical disease such as cervical cancer and viral hepatitis patients. This investigation has some significance to Instruct clinical diagnosis, differential diagnosis and treatments.