液相等电聚焦电泳纯化燕窝蛋白质

Purification of Proteins from Edible Bird s Nest by Liquid-Phase Isoelectric Focusing

以市售燕窝为材料,采用MicroRotofor系统进行液相等电聚焦电泳(liquid-phase isoelectric focusing,LIEF)纯化燕窝蛋白质,等电聚焦电泳后对收集到10个小室的样品进行SDS凝胶电泳分析。结果显示,燕窝蛋白质提取液中的一些胶状物聚集在聚焦槽酸端第1～4样品室,第9、10样品室杂质也较多,合并第5～8样品室的样品,即得纯化后的燕窝蛋白用于后续双向电泳分析。液相等电聚焦电泳预分离纯化技术结合双向电泳技术,对4个燕窝蛋白质进行较好的分离,在2-DE胶上检测到约30～60个蛋白质点。由此可见,液相等电聚焦电泳预分离技术是燕窝蛋白质进行2-DE分离前的重要纯化手段。

Liquid-phase isoelectric focusing（LIEF） was applied to purify proteins extracted from edible bird s nest（EBN）.After LIEF prefractionation,totally 10 fractions were collected and analyzed with SDS-polyacrylamide gel electrophoresis（PAGE）.The results showed that gelationus materials mainly concentrated at the first four fractions,and there were also some impurities at the last two fractions.So fractions five through eight were pooled as purified protein sample for following two-dimensional electrophoresis（2-DE） analysis.By using LIEF combined with 2-DE,4 proteins from EBNs were well separated,and about 30 to 60 2-DE spots were detected on the 2-DE gels.The findings confirmed that LIEF as a prefractionation prior to IPG-IEF can produce the best 2-DE patterns for EBN.