添加剂对喷雾干燥干扰素α-2b稳定机制的研究

Mechanism of Recombinant Human Interferon α-2b Stabilization by Additives During Spray-Drying

目的研究添加剂对蛋白质在喷雾干燥过程中的稳定机制。方法以乳糖或甘露醇作为赋形剂,并以聚山梨酯20、十二烷基硫酸钠、L-赖氨酸、羟丙基甲基纤维素或羟丙基-β-环糊精为添加剂将干扰素α-2b制成喷雾干燥粉末。用X光电子能谱分析研究干扰素α-2b的表面吸附现象,用圆二色谱测定该蛋白的二级结构。结果上述几种添加剂均能降低干扰素的表面吸附量;当以乳糖(而非甘露醇)为赋形剂时,添加剂的加入提高了该蛋白α螺旋结构的含量。结论蛋白质在喷雾干燥过程中的稳定性受载体结晶、表面富集和构象变化等多种机制影响。

OBJECTIVE To investigate the mechanism of protein stabilization by additives during spray-drying, METHODS Recombinant human interferon α-2b （IFN α-2b） was spray dried, with lactose or mannitol as vehicles and Tween 20, SDS, L-lysine, HPMC and HP-β-CD as additives. X-ray photoelectron spectroscopic analyses were performed to investigate the surface adsorption of IFN α-2b, and the secondary structure of the protein in these formulations was characterized using circular diehroism. RESULTS All of the additives decreased the amount of IFN α-2b on the surface. The inclusion of the additives raised the α-helix content when lactose, but not mannitol, was used as vehicles. CONCLUSION The stability of the protein during spray drying was affected by crystallizalion, surface adsorption and conformation transformation.