疏水性电荷诱导层析纯化免疫球蛋白IgY

Purification of immunoglobulin IgY with hydrophobic charge induction chromatography

利用疏水性电荷诱导层析(HCIC)从鸡血中分离免疫球蛋白IgY。采用经辛酸预处理后的血浆上清液作为原料,直接用HCIC分离纯化IgY。考察了两种HCIC介质,市售介质MEP HyperCel和实验室自制介质Bestarose-DVS-MMI,优化了上样pH、洗脱pH和上样量等条件。研究发现,以2-巯基-1-甲基咪唑为配基的Bestarose-DVS-MMI介质优于MEP HyperCel介质,前者分离IgY的纯度和收率较高,HPLC分析IgY纯度可达92%以上,收率约95%。通过配基结构比较和表面电势分析,探讨了HCIC分离IgY的机制。结果表明,采用HCIC可以实现IgY的高效分离,为从血液中分离免疫球蛋白提供了新方法。

Hydrophobic charge induction chromatography（HCIC）was used to separate immunoglobulin IgY from chicken plasma.The plasma was pretreated with caprylic acid to precipitate some impurities,and the supernatant was used directly as the feedstock for separation and purification of immunoglobulin IgY by HCIC.Two resins,commercial MEP HyperCel and home-made Bestarose-DVS-MMI,were used as HCIC medium and investigated.Operation conditions were optimized,including pH value of sample loading and elution,as well as sample loading volume.It was found that Bestarose-DVS-MMI with 2-mercapto-1-methyl-imidazole as the ligand was better than MEP HyperCel.The results indicated that IgY could be separated with high purity（above 92% with HPLC analysis）and yield about 95%.The mechanism of HCIC was discussed based on comparison of ligand structure and surface potential.The results demonstrated that HCIC can be efficiently used for IgY purification from chicken plasma,which can be a new approach for the immunoglobulin separation from the plasma.