Preparation And Labeling Of Antibody Against 3α-Hydroxysteroid Dehydrogenase

Comamonas testosteroni containing steroids degradation of key 3α-Hydroxysteroid dehydrogenase(3α-HSD) gene,can completely absorb this type of substrate through a series of complex metabolic way.After immunization of BALB/c mice with purified 3α-Hydroxysteroid dehydrogenase(3α-HSD),three hybridoma cell strains against 3α-HSD were developed after fusion between SP2/0 myeloma cell and the stimulated splenocytes.The indirect ELISA results showed that the McAbs ascites titer were 1×10~6,1×10~5,1×10~4 separately.The three McAbs were purified by protein G-Sepharose and labelled with HRP and FITC separately.The results of test showed that the enzyme-labeled antibody and the fluorescent antibody have highly specificity and sensitivity.Application of the technology of monoclonal antibodies for the monitoring of water pollution was detected.The results showed that the monoclonal antibody has high specificity for steroids.