摘要
为分离纯化纳豆激酶,使用纳豆激酶粗制液经硫酸铵分级盐析、Sephodex G100和CM-Sephrose-6B FF层析等步骤纯化,经SDS-聚丙烯酰胺凝胶电泳检测。以纯化的纳豆激酶作为抗原,结合福氏佐剂,采用多次多点肌肉注射法对兔进行免疫,以琼脂免疫双向扩散法检测效价。试验结果表明,纳豆激酶粗制液经纯化后,得纯酶样品,酶被纯化了28.4倍,比活力为608.6 Uku.mg-1蛋白,SDS-聚丙烯酰胺凝胶电泳检测为单一蛋白条带,纳豆激酶兔抗血清效价为1∶64。成功制备的兔抗纳豆激酶抗血清,为研究纳豆激酶的功能提供了物质基础。
To separate and purify the Nattokinase and to prepare antiserum against Nattokinase. Nattokinase was purified through procedures including ammonium sulfate differential precipitation, sephadexG-100, and CM-Sephrose-6B FF chromatography, and was detected on gels of SDS-PAGE. Rabbit antiserum against Nattokinase was prepared by muscle immunization mixed with Freund＇s adjuvant and the titer was detected by double immunodiffusion carried out on agar gel. The purified sample＇s specific activity was 608.6 UKu·mg^-1 protein and was purified by 28. 4 fold. The purified nattokinase showed single protein band on gels of SDS-PAGE. The titer of rabbit antiserum against Nattokinase was 1 ： 64. The rabbit antiserum against Nattokinase is prepared successfully, and it provides the material foundation to the function study of Nattokinase.
出处
《山西农业大学学报（自然科学版）》
CAS
2009年第4期308-311,共4页
Journal of Shanxi Agricultural University(Natural Science Edition)
基金
山西省青年科学基金(20041036)
关键词
纳豆激酶
分离纯化
抗血清
Nattokinase separation and purification
antiserum
