摘要
本实验研究了离子交换色谱技术大规模纯化乳铁蛋白的色谱条件。选用的SP Sepharose Fast Flow离子交换剂对乳铁蛋白有很好的吸附选择性,洗脱速度可达到2L/h。以pH值为7.7～8.0,10mmol/L的磷酸盐为起始缓冲液,通过改变离子强度进行分步洗脱。先采用0.5mol/L的NaCl溶液选择性洗脱分泌型免疫球蛋白A和乳过氧化物酶组分,再采用1mol/L的NaCl洗脱LF组分。经SDS-PAGE测定,所得乳铁蛋白组分显示为单一区带,相对分子质量为80400Da。经等点聚焦测定,所得乳铁蛋白的等电点为8.65。中试生产的精制品中乳铁蛋白的纯度和回收率分别为94.20%和75.45%。
A stepwise procedure for purification of lactoferrin （LF） extracted from bovine colostrum by ion exchange chromatography on a pilot scale was investigated. SP Sepharose Fast Flow （SP Sepharose FF） of excellent absorption speciality for LF, was chosen as the ion exchanger with elution rate of 2 L/h. 0.5 mol/L NaC1 aqueous solution was used to elute the secretory immunoglobulin A and lactoperoxidase. Then, lactofen＇in was eluted with 1.0 mol/L NaCl aqueous buffer. Lactoferdn fraction is shown as a single band in SDS-PAGE with molecular weight of 80400 Da. The isoelectdc point of lactoferrin is 8.65 determined by isoelectric focusing. The purity of refined LF on pilot production is 94.20% with a yield of 75.45%.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2007年第7期48-53,共6页
Food Science
基金
江南大学校预研项目(2004LYY028)
关键词
乳铁蛋白
分离纯化
强阳离子交换色谱
中试规模
lactoferrin
purification
strong cation exchange chromatography
pilot scale
