title: [Isolation and purification of recombinant soluble and non-fusion angiogenesis inhibitor Kringle 5 using chromatography].
abstract: The Kringle 5 domain of plasminogen is one of the most potent angiogenesis inhibitors known to date, which can inhibit cell proliferation and migration efficiently. In the study, on the foundation of successful clone and expression of recombinant soluble and non-fusion angiogenesis inhibitor Kringle 5, a two-step chromatographic method, including the use of SP Sepharose Fast Flow cation exchanger and Sephacryl S-100 HR size exclusion chromatography in sequence, was established to separate and purify angiogenesis inhibitor Kringle 5. On the SP Sepharose Fast Flow column, the buffer A consisted of 50.0 mmol/L acetic acid-sodium acetate (pH 5.2), and the buffer B consisted of buffer A with the addition of 0.5 mol/L sodium chloride (pH 5.2); on Sephacryl S-100 HR column, the elution buffer was 5.0 mmol/L phosphate solution (pH 7.0). Through the two-step chromatographic purification process, the purity of the obtained Kringle 5 was more than 98%. In addition, it was found that the obtained Kringle 5 could inhibit the blood vessel growth of chick embryo chorioallantoic membrane effectively. Finally it is concluded that this method can effectively separate active recombinant soluble and non-fusion angiogenesis inhibitor Kringle 5.
authors.affiliation: College of Life Science, Northwest University, Xi' an 710069, China.
authors.forename: Lina
authors.initials: L
authors.lastname: Ma
authors.forename: Dan
authors.initials: D
authors.lastname: Wu
authors.forename: Liujiao
authors.initials: L
authors.lastname: Bian
mesh_terms: D020533:Angiogenesis Inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D002411:Cation Exchange Resins / Q000737:chemistry; D002412:Cations / Q000737:chemistry; D002850:Chromatography, Gel / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D010446:Peptide Fragments / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D010958:Plasminogen / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D012995:Solubility
chemical_list: D020533:Angiogenesis Inhibitors; D002411:Cation Exchange Resins; D002412:Cations; D010446:Peptide Fragments; D011994:Recombinant Proteins; C491499:plasminogen kringle 5; D010958:Plasminogen