多壁碳纳米管固相萃取-在线提取蛋清中的溶菌酶

On-line Isolation of Lysozyme from Egg-white Using Multi-walled Carbon Nanotubes

建立了以羧基功能化的多壁碳纳米管作为吸附剂在线提取蛋清中溶菌酶的新方法.将多壁碳纳米管经氧化和提纯后装入固相萃取柱,在顺序注射系统中实现对蛋清中溶菌酶的在线选择性萃取,以碳酸盐缓冲溶液定量洗脱,进样量为2 mL时,富集倍率为12,吸附率和洗脱率均为100%,采样速率为10 h-1,精密度为3.0%.实验结果表明,该方法所用试剂量少,提取速度快,每100 mL蛋清可提取溶菌酶0.4 g,且溶菌酶纯度较高.

A novel procedure for the extraction of lysozyme from egg-white using carbon nanotubes as a sorbent was developed. Carbon nanotubes after oxidation and purification were used to pack a micro-column which was incorporated into a sequential injection system for facilitating online selective sorption of lysozyme from egg-white. The retained protein was afterwards quantitatively eluted by a carbonate buffer. With a sample loading volume of 2.0 mL, an enrichment factor of 12 was obtained, along with a retention efficiency of 100% and a recovery rate of 100%. A sampling frequency of 10 h^-1 was achieved along with a precision of 3.0% RSD. Lysozyme of 0.4 g could be extracted from 100 mL egg-white. SDS-PAGE demonstrated that the purity of the obtained lysozyme was as high as that of pure reagent.