Isolation And Purification Of Recombinant Human Stromal Cell-derived Factor1α With Column ChromatograpHy

Vascular endothelial growth affects the pHysiological and pathological process of many diseases, such as clinically detectable tumor growth and metastasis.The cardiovascular and cerebrovascular diseases are dependent on angiogenesis too. At present, people have developed a lot of angiogenic factors can directly stimulate the blood vessels, such as VEGF, FGF, Angiopoietin-I and Angiogenin, but there still have quite a lot of proplems like the nitric oxide mediated hypotension, vascular malformation and increased vascular permeability, which can not be carried out on their drug development or application research. And stromal cell derived factor1alpHa (stromal cell-derived factor-1a, SDF-1a) can promote the mobilization of stem cells in vivo and induce angiogenesis. They are so far found to be the strongest chemokines that promote the mobilization of stem cells. Nevertheless SDF-la does not directlypromote angiogenesis,however it can gather multiple cell angiogenesis in ischemic. Therefore, SDF-1is a drug for the treatment of myocardial ischemia, the potential of cerebral ischemic and ulcer diseas.This work demonstrated the isolation and purification of the refolded recombinant human stromal cell-derived factor1a (SDF-1a) through a two-step chromatograpHic method consisting of SP-Sepharose Fast Flow cation exchange chromatograpHy Sephadex G-75size-exclusion chromatograpHy. SDS-PAGE and high-performance chromatograpHic analysis showed that the derived SDF-la had a purity of more than96%; Cell chemotaxis of SDF-la on monocyte THP-1showed that the resulting SDF-la induces chemotactic activity on THP-1cell. After the two-step purification by column chromatograpHy, the purity of SDF-la had increased by about160times, which approximates a14%yield.