Expression and purification of recombinant ATF-mellitin, a new type fusion protein targeting ovarian cancer cells, in XXXX .

@entity1472 @entity137 @entity5 @entity10056 @entity1042 @entity3 @entity214
UNASSIGNED: Melittin is well known to possess cytolytic activity with wide-spectrum lytic properties and its potential use as an agent to treat several types of @entity5 has been tested. Due to the non-specific @entity137 , melittin can impair not only @entity5 cells but also normal tissue. Thus, @entity5 -targeted toxins may be helpful for developing novel anticancer therapeutics. The urokinase-type plasminogen activator (uPA) plays a central role in tissue remodelling events occurring in normal physiology and in pathophysiology, including @entity3 and @entity3 . Heartening findings showed that uPA receptor is predominantly expressed on many types of @entity5 . Therefore, the @entity1472 -terminal fragment (ATF) of uPA which was able to identify and bond with @entity5 cells was used as the cell-targeting domain to make up @entity5 -targeted toxin in this study. In the present study, pPICZaC-ATF-melittin eukaryotic expression vector was successfully constructed. After transformed into @entity10056 and induced by @entity1042 , rATF-mellitin was detected by @entity214 -PAGE and western blot analysis. After induction with @entity1042 , the expression level of rATF-mellitin was 312  mg/l in 80-l fermentor. rATF  -mellitin was purified to >95% purity using SP Sepharose ion exchange chromatography and source    30  RPC with 67.2% recovery. Cell proliferation assay showed that rATF-melittin inhibited growth of SKOV3 cells and had no @entity137 effect on normal cells. For the first time, we established a stable and effective rATF-mellitin @entity10056 expression system to obtain a high level of expression of secreted rATF-mellitin which was purified by a highly efficient purification procedure.
