Abstract :
In this study, chemiluminescence immunoassay was used to screen clones of mouse anti-human albumin monoclonal antibody (anti-ALB McAb). The method is simplified as a one-step operation and the experimental parameters are optimized by orthogonal experiment [L9 (34)]. The parameters of the positive clone / blank control luminescence value (SNR) are the largest and the method is evaluated and statistical analysis. The results showed that the coated antibody was 3mg / L, the ratio of the enzyme protein A was 1: 2000, the incubation time was 304 times and the signal - to - noise ratio was 1284, which was several times higher than other combinations. The linear range was 20-20000ng / L, the precision was good, the average CV was 5.32% and the average inter-assay CV was 8.82%. The results showed that the method can be used for cloning and screening of anti-ALB McAb and the method is obviously superior to ELISA and other traditional methods, especially the simple and rapid operation of one step is also suitable for high antibody screening of other antibodies.
Keyword :
chemiluminescence immunoassay, anti-human albumin monoclonal antibody, orthogonal test, clone screening