食品科学 ›› 2008, Vol. 29 ›› Issue (4): 87-89.

• 基础研究 • 上一篇    下一篇

DEAE-52层析对猪血清IgG提纯得率的影响

刘生杰, 余为一, 朱茂英, 刘成文, 聂传朋, 李东伟   

  1. 阜阳师范学院生物系; 安徽农业大学生命科学学院; 阜阳市畜牧局; 阜阳师范学院生物系
  • 出版日期:2008-04-15 发布日期:2011-08-24

Effects of DEAE-52 Anion-exchanging Chromatography on Yield Rate of IgG Purified from Swine Serum

 LIU  Sheng-Jie, YU  Wei-Yi, ZHU  Mao-Ying, LIU  Cheng-Wen, NIE  Chuan-Peng, LI  Dong-Wei   

  1. 1.Department of Biology, Fuyang Teachers College; 2.School of Life Science, Anhui Agricultural University; 3.Husbandry Bureau of Fuyang
  • Online:2008-04-15 Published:2011-08-24

摘要: 目的:提纯猪血清IgG,并分析DEAE-52层析对IgG得率的影响。方法:用硫酸铵盐析和DEAE-52阴离子交换层析两步法提纯猪血清IgG,以SDS-PAGE、Bradford法浓度测定和免疫双扩散来分析DEAE-52层析对IgG得率的影响。结果:DEAE-52层析获得两个分开明显无重叠洗脱峰,两峰均含有IgG,其中第一峰蛋白纯度达95.7%,得率为3.0~4.0mg/ml血清;第二峰IgG纯度59%。结论:DEAE-52层析能使猪血清IgG在两个洗脱峰中分布,两步法能从层析第一峰获得电泳纯IgG,对层析第二峰进一步纯化能大幅提高IgG得率。

关键词: 猪IgG, 提纯, 层析, 得率, 影响

Abstract: Objectives: To purify immunoglobulin G (IgG) from swine serum and analyze the effects of DEAE-52 chromatog- raphy on the yield rate of IgG. Methods: Porcine IgG was separated and purified from swine serum by two-step methods, SDS- PAGE and Bradfold assaying kit with saturated ammonium sulphate (SAS) precipitation followed by DEAE-52 cellulose anion- exchanging chromatography. The effects of DEAE-52 chromatography on the yield rate of IgG were defermined by SDS-PAGE and Bradford assaying kit with double immunodiffusion(DI). Results:Two different eluate peaks are obtained in DEAE-52 chromatography. IgG is also found in different eluate peaks of chromatogram by SDS-PAGE and DI.The yield rate and purity of IgG in first eluate peak are 3.0~4.0 mg/ml serum and 95.7% respectively. But the purity in second eluate peak is lower. Conclusion: DEAE-52 chromatography leads the porcine IgG to distribute in two different eluate perks. The IgG of electro- phoretic purity can be purified from the first eluate perk of DEAE-52 chromatogram by two-step methods. The yield rate of IgG can be largely increased by IgG and further purified from the second eluate perk .

Key words: porcine immunoglobulin G(IgG), purify, chromatography, yield, effect