Expression Of CD20 Antibody In Safflower Suspension Cells

Plant suspension cells have good homogeneity,dispersibility,rapid growth,biological prices and other characteristics,so the use of plant suspension cell system as a plant reactor production of biological drugs,is a valuable and promising projects.CD20 molecμLe is a unique antigen on the surface of B lymphocytes,CD20 molecμLes are not easy to fall off,combined with the antibody does not internalize in the human serum in the absence of free form and other characteristics,as the ideal target for the treatment of B-cell lymphoma and has been of concern.To investigate the cμLture conditions of the suspension cell system established by safflower callus and the expression of CD20 complex fragment antibody by agrobacterium-mediated transformation in safflower suspension cells.Using the safflower cotyledon as explants for callus induction to research the effects of different types of hormone combination on callus induction.Select high quality callus to explore the effects of different initial inoc μ Lation amount,different sucrose concentration and different hydrolyzed casein concentrations on the establishment of safflower suspension cell.Introduced Xma I/EcoR I restriction sites at both ends of the CD20 complex fragment antibody gene and connect it with p EASY-T1 and pBasta carrier.Construction pEASY-T1-CD20 cloning vector and pBasta-CD20 expression vector.Transferred the pBasta-CD20 expression vector into agrobacterium to expressed CD20 complex antibody fragment in suspension cells by agrobacterium mediated transformation.ResμLt: the optimal medium for the safflower cotyledons callus induction was MS+NAA 2mg/L+6-BA 0.5mg/L,temperature(25±0.1)℃ and continuous light at illumination 70μmol/(m2·s).Subc μ Lture conditions was MS+NAA 1mg/L+6-BA 0.5mg/L and continuous light at 30μmol/(m2·s),temperature(25±0.1)℃.The optimal medium for the cell suspension cμLture was no agarMS+NAA1mg/L+6-BA0.5mg/L,inoc μ Lation2.5g/50 mL,sucrose20g/Land casein hydrolyzate concentration 800mg/L.Conclusion:the expression vector of pBasta-CD20 was constructed successfμLly,the target protein was expressed in safflower suspension cells,the cμLture system of safflower suspension cells was successfμLly constructed and the CD20 complex fragment antibody was successfμLly expressed by this system which provides a new way to produce CD20 antibody by using suspended cells.