Qiu P, Ding Y, Qin J, Han K K, Zhu D
Department of Biochemistry, Nanjing University, P.R. China.
Biol Chem Hoppe Seyler. 1994 Jun;375(6):413-8. doi: 10.1515/bchm3.1994.375.6.413.
The human macrophage colony stimulating factor (hM-CSF) in its monomeric form has been over-produced in BmN cells and in silkworm larvae infected with the recombinant baculovirus Bm284M-CSF. The recombinant monomeric M-CSF (rhM-CSF) exhibited the activity of 8-14 x 10(4) units/ml of cell culture medium. When the insect larvae were infected with the recombinant virus, the maximum rhM-CSF was expressed 4-5 days post infection with an activity of 3 x 10(6) units/ml hemolymph. The monomeric rhM-CSF was purified to homogeneity through three steps of purification. A pilot purification yielded 1 mg of homogeneous monomeric rhM-CSF from 10 larvae. The purified rhM-CSF monomers gradually dimerized in vitro. In contrast, the crude or the semi-purified monomers did not dimerize in vitro, indicating that the presence of an unknown moiety in the rhM-CSF preparations obtained from hemolymph interfered with dimerization.
人巨噬细胞集落刺激因子（hM-CSF）的单体形式已在BmN细胞和感染重组杆状病毒Bm284M-CSF的家蚕幼虫中过量产生。重组单体M-CSF（rhM-CSF）在细胞培养基中的活性为8-14×10⁴单位/毫升。当昆虫幼虫感染重组病毒时，感染后4-5天rhM-CSF表达量最高，血淋巴中的活性为3×10⁶单位/毫升。单体rhM-CSF通过三步纯化达到均一性。小规模纯化从10只幼虫中获得了1毫克均一的单体rhM-CSF。纯化后的rhM-CSF单体在体外逐渐二聚化。相比之下，粗提物或半纯化的单体在体外不会二聚化，这表明从血淋巴中获得的rhM-CSF制剂中存在未知成分会干扰二聚化。
