[Two-step chromatographic method for the separation and purification of recombinant angiogenesis inhibitor Kringle 5].
The Kringle 5 domain of plasminogen, which was previously shown to inhibit angiogenesis in vitro and in vivo, is one of the most potent angiogenesis inhibitors known to date. A two-step chromatographic method, which consists of Chelating Sepharose Fast Flow chelated with Ni2+ affinity medium and Sephadex G-75 medium, was established to separate and purify recombinant angiogenesis inhibitor Kringle 5 (rK5). Through the two-step chromatographic purification process, the obtained rK5 was confirmed to be homogeneous on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and its relative molecular mass was estimated to be about 32 000, which matched well with the prediction by gene sequence. Its purity was about 98%, and the total protein recovery of this method was 0.63%. In addition, it was found that it inhibited the blood vessel growth of chick embryo chorioallantoic membrane effectively.
National Engineering Research Center for Miniaturized Detection System, College of Life Science, Northwest University, Xi'an 710069, China.
Xingui
X
Jia
Liujiao
L
Bian