Abstract:
ObjectiveTo construct and express the single chain Fv gene fragment for human CD
3 antigen and conduct imitation analysis on conformation.
MethodsThe variable region gene of heavy chain(VH)and light chain(VK)of antiCD
3 antibody were linked into single chain Fv by splicing overlapping and extension(SOE)ScFv genes were cloned to pET28(a+)and expressed in E.coli.
ResultsThe DNA sequence analyses and imitation of conformration analyses demonstrated that ScFv gene were 732bp,conformation steady.compatibility S>0.The ScFv fragment were expressed in BL21(DE3),and folded into 26.5 KD protein.
ConclusionThe correct gene of anti-CD
3 Scfv was constructed.It would be used to construct the anti-CD
3/anti-HBV bifunctional antibody.