CAJ | 학술논문

干细胞因子是一种多功能细胞因子,能在多级造血水平与其他细胞因子协同作用促进造血干/祖细胞及各种血细胞的存活、增殖和分化.重组人二连体干细胞因子具有比干细胞因子单体更高的比活,可以避免副反应.重组人二连体干细胞因子在Sf9细胞和大肠杆菌中表达时,发现有特异性降解.片段缺失实验证实切割位点位于重组人二连体干细胞因子的145位到165位氨基酸之间.丝氨酸蛋白酶抑制剂aprotinin和PMSF能抑制这种特异性降解.试验了不同浓度的aprotinin和PMSF对Sf9细胞存活和重组人二连体干细胞因子产量的影响,显示当aprotinin的浓度为1.0μg/mL时,重组人二连体干细胞因子的产量是没有加aprotinin时产量的2倍,而且aprotinin可以完全抑制这种特异性降解.
간세포인자시일충다공능세포인자,능재다급조혈수평여기타세포인자협동작용촉진조혈간/조세포급각충혈세포적존활、증식화분화.중조인이련체간세포인자구유비간세포인자단체경고적비활,가이피면부반응.중조인이련체간세포인자재Sf9세포화대장간균중표체시,발현유특이성강해.편단결실실험증실절할위점위우중조인이련체간세포인자적145위도165위안기산지간.사안산단백매억제제aprotinin화PMSF능억제저충특이성강해.시험료불동농도적aprotinin화PMSF대Sf9세포존활화중조인이련체간세포인자산량적영향,현시당aprotinin적농도위1.0μg/mL시,중조인이련체간세포인자적산량시몰유가aprotinin시산량적2배,이차aprotinin가이완전억제저충특이성강해.
Stem cell factor (SCF) is a hematopoietic cytokine that promotes the survival,proliferation, and differentiation of hematopoietic cells. The recombinant dual human stem cell factor (rdhSCF) possesses higher specific activity than the monomer form of hSCF and might be preferred since it could be administered at low doses to avoid significant mast cell activation while stimulating hematopoietic recovery. In this report, degradation was found when rdhSCF was expressed in Sf9 cells and E. coli cells. The deletion analysis of rdhSCF confirmed that the cleavage site locates in the region from 145 to 165 amino acid of rdhSCF. Only serine protease inhibitor aprotinin and PMSF could inhibit the site-specific degradation. The influences of aprotinin and PMSF on Sf9 cell survivals and rdhSCF yield were measured at different concentrations. When aprotinin concentration was 1.0 μg/mL, the highest expression level of rdhSCF was achieved, which was about two times higher than without adding protease inhibitor. Aprotinin could completely inhibit degradation of rdhSCF expressed in insect cells.