Moshashaée S, Bisrat M, Forbes R T, Nyqvist H, York P
AstraZeneca, Södertälje, 151 85, Södertälje, Sweden.
Eur J Pharm Sci. 2000 Sep;11(3):239-45. doi: 10.1016/s0928-0987(00)00108-1.
The solution enhanced dispersion by supercritical fluid (SEDS) process was used to evaluate the effect of the processing variables on the biological and physicochemical characteristics of lysozyme protein particles produced from an organic solution of dimethylsulfoxide (DMSO) using an experimental design procedure. The processing variables were temperature, pressure, solution concentration and the flow-rates of supercritical carbon dioxide and a protein solution. Solutions of hen egg lysozyme (0.5-1%, w/v) in DMSO were dispersed using supercritical carbon dioxide as the antisolvent, and particles precipitated in a particle formation vessel. The morphology, particle size and size distribution and biological activity of the protein were determined. The precipitates were also examined with high sensitivity differential scanning calorimetry (HSDSC) and high-performance cation-exchange chromatography. The amount of residual DMSO was determined using headspace gas chromatography. Particle size measurements showed the precipitates to be agglomerates with primary particles of size 1-5 microm, containing <20 ppm of residual solvent. The activity of the precipitates varied between 44 and 100% depending on the experimental conditions. The similarity of HSDSC data for unprocessed and processed samples indicated that the SEDS process does not cause major denaturation of lysozyme when prepared from DMSO solutions. By optimising of working conditions, the SEDS process can produce micron-sized particles of lysozyme with minimal loss of biological activity.
采用超临界流体增强分散法（SEDS），通过实验设计程序评估工艺变量对由二甲基亚砜（DMSO）有机溶液制备的溶菌酶蛋白颗粒的生物学和物理化学特性的影响。工艺变量包括温度、压力、溶液浓度以及超临界二氧化碳和蛋白质溶液的流速。以超临界二氧化碳作为抗溶剂，将蛋清溶菌酶（0.5 - 1%，w/v）在DMSO中的溶液进行分散，颗粒在颗粒形成容器中沉淀。测定了蛋白质的形态、粒径和粒径分布以及生物学活性。还用高灵敏度差示扫描量热法（HSDSC）和高效阳离子交换色谱法对沉淀物进行了检测。使用顶空气相色谱法测定残留DMSO的量。粒径测量表明沉淀物为团聚体，初级颗粒尺寸为1 - 5微米，残留溶剂含量<20 ppm。沉淀物的活性根据实验条件在44%至100%之间变化。未处理和处理后样品的HSDSC数据相似，表明从DMSO溶液制备溶菌酶时，SEDS工艺不会导致溶菌酶发生重大变性。通过优化工作条件，SEDS工艺可以生产出具有最小生物活性损失的微米级溶菌酶颗粒。
