Soluble multimer of recombinant endostatin expressed in E. coli has anti-angiogenesis activity.

Abstract:

:The bioactivity, refolding, and multimer formation of endostatin, particularly of recombinant endostatin produced from bacteria, are proved challenging for clinical application. In order to determine the biological activity of recombinant endostatin multimer, first, we expressed endostatin in Escherichia coli and purified it with ion-exchange chromatography. The purified active protein could elicit multimer formation spontaneously, but still has comparable activity. Aim to determine the anti-angiogenic activity of multimer endostatin, by use of RP-HPLC, we then successfully separated endostatin monomer and multimer for subjecting to anti-angiogenesis assay. The results from CAM (chorioallantoic membrane) inhibition assay showed that both monomer and multimer suppressed CAM vascularization significantly. At the dosage of 0.8 microg, inhibition rates of multimeric and monomeric proteins were about 58% and 38%, respectively. Multimeric endostatin exerted a higher activity than monomeric endostatin (p < 0.05). However, when the protein dosage is less than 0.4 microg/ml, there is no significance between their inhibition rates (p > 0.05), although both of them show a high inhibition effect in contrast to control. The results from HUVEC proliferation assay also showed similar effects at dosages of 0.6 and 1.6 microg/ml, multimer exerted a higher activity on inhibition of HUVEC proliferation comparing with monomer (p < 0.05). In conclusion, our results suggest that endostatin multimer has a comparable or higher bioactivity and multimerization will not affect its bioactivity, implying that endostatin activity is insensitive to structure conformation contributed by disulfide bonds.

authors

Wei DM,Gao Y,Cao XR,Zhu NC,Liang JF,Xie WP,Zhen MY,Zhu MS

doi

10.1016/j.bbrc.2006.05.031

subject

Has Abstract

pub_date

2006-07-14 00:00:00

pages

1398-404

issue

4

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(06)01072-2

journal_volume

345

pub_type

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