Abstract: Combining hydrophobic charge-induction chromatography (HCIC) and expanded bed adsorption (EBA), a novel efficient bioseparation process was developed to purify the immunoglobulin IgY from poultry blood. New hydrophobic charge-induction expanded bed adsorbent S-MMI was prepared with 2-mercapto-1-methyl imidazole (MMI) as the HCIC ligands coupled to quartz-densified agarose matrix for EBA. S-MMI had high ligand density (about 105 mmol·g^-1) and the saturated adsorption capacity for IgY reached 71.26 mg·ml^-1 at pH 7. S-MMI could form perfectly classified fluidization in expanded bed with high stability. Chicken plasma was precipitated with caprylic acid and the supernatant was used as the feedstock for IgY separation. The loading and elution conditions were optimized by packed bed chromatography. The effects of expansion factors in expanded bed were investigated and the separation conditions were optimized. Finally, IgY was purified from pretreated chicken plasma by EBA with high purity of 98.9% and yield of 80.5%. The combination of HCIC and EBA could achieve efficient separation of immunoglobulin from crude feedstock, which provided a new way for the comprehensive utilization of poultry blood resource.

Key words: bioseparation, adsorption, chromatography, expanded bed, hydrophobic charge-induction, IgY, chicken blood

摘要： 结合疏水性电荷诱导色谱和扩张床吸附两项新型生物分离技术，开发了一个高效分离禽类血液免疫球蛋白IgY的新过程。以2-巯基-1-甲基咪唑（MMI）为疏水性电荷诱导配基，偶联于琼脂糖扩张床吸附基质上，制备出疏水性电荷诱导扩张床吸附剂S-MMI。结果表明，S-MMI具有较高的配基密度，约105 mmol·g^-1，pH7时IgY饱和吸附容量达71.26 mg·（ml介质）^-1，扩张床内可形成稳定的分级分布，床层稳定。选用经辛酸沉淀预处理后鸡血浆为料液，通过固定床色谱确定了上样和洗脱条件，比较了扩张床中不同膨胀率的影响，优化了分离条件，实现了扩张床吸附分离鸡血IgY，纯度达到98.9%，收率为80.5%。结果表明，疏水性电荷诱导色谱结合扩张床吸附，可以实现免疫球蛋白的高效分离，为禽类血液蛋白综合利用提供了新方法

关键词: 生物分离, 吸附, 色谱, 扩张床, 疏水性电荷诱导, IgY, 鸡血