Production Of Monoclonal Antibody To Lipopolysaccharide And C99

Monoclonal antibody technology is the important tool of modern life science research and have an indispensable role in the study of the structure and function of genes and protein.This article mainly chooses lipopolysaccharide(LPS)and C99 as immunogen for Balb/c mice and use classic hybridoma technology to establish a stable hybridoma cell lines of secreting specificity monoclonal antibody.We expand the culture of hybridoma cell lines and purifyto obtain the monoclonal antibodies.It is applied to the research of cell of disease model and provide the raw materials for disease prevention and control.In the meantime,it may also be used to study the diagnosis and prevention of the disease.When animal serum titer reaches a stable level,we choose the animal of owing best serum titer by ELISA to operate cell fusion experiment and implant cell into 96-well cell culture plates,cell culture plate into the 96 holes in cell culture.After the culture of a period of time,we operate the screening of positive hybridoma cell lines and obtain respectively10 and 8 positive hybridoma cell lines for LPS and C99.In the meantime,we eliminate the false positive hybridoma cell lines through the second screening.Then,we choose 3hybridoma cell lines having the best titer by to operate cell subclone experiment and obtain monoclonal cell lines.Their serial numbers respectively are 1B9-H7-H6,5C12-A12-C8,5F6-G8-B11 and 1H5-A10-H7,3H2-A9-D9 for LPS and C99.We shift the cell into cell culture bottles to expand the cell culture size after obtaining monoclonal cell lines for LPS and C99 monoclonal cell lines.In the meantime,we can also inject cells into enterocoelia of mouse to collect antibody,then we can achieve the batch production of monoclonal antibodies through these methods.Finally,we identificate the subclass of monoclonal antibodies through the mouse monoclonal antibody subclass testing kits and get the result that the subclass of these antibodies is IgG2 A,IgG2B and IgM.