疏水性电荷诱导扩张床吸附分离免疫球蛋白IgY

doi:10.3969/j.issn.0438-1157.2014.01.025

化工学报 ›› 2014, Vol. 65 ›› Issue (1): 198-204.DOI: 10.3969/j.issn.0438-1157.2014.01.025

疏水性电荷诱导扩张床吸附分离免疫球蛋白IgY

施伟, 林东强, 姚善泾

化学工程联合国家重点实验室, 浙江大学化学工程与生物工程学系, 浙江杭州 310027

收稿日期:2013-08-19 修回日期:2013-10-16 出版日期:2014-01-05 发布日期:2014-01-05
通讯作者: 林东强
作者简介:施伟（1986-），男，博士研究生。
基金资助:
国家自然科学基金项目；高等学校博士学科点专项科研基金项目；浙江省自然科学基金项目。

Separation and purification of immunoglobulin IgY with hydrophobic charge-induction expanded bed adsorption

SHI Wei, LIN Dongqiang, YAO Shanjing

State Key Laboratory of Chemical Engineering, Department of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, Zhejiang, China

Received:2013-08-19 Revised:2013-10-16 Online:2014-01-05 Published:2014-01-05
Supported by:
supported by the National Natural Science Foundation of China, Specialized Research Fund for the Doctoral Program of Higher Education and the Natural Science Foundation of Zhejiang Province.

摘要/Abstract

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

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^-1at 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

中图分类号:

TQ028.8

施伟, 林东强, 姚善泾. 疏水性电荷诱导扩张床吸附分离免疫球蛋白IgY[J]. 化工学报, 2014, 65(1): 198-204.

SHI Wei, LIN Dongqiang, YAO Shanjing. Separation and purification of immunoglobulin IgY with hydrophobic charge-induction expanded bed adsorption[J]. CIESC Journal, 2014, 65(1): 198-204.

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疏水性电荷诱导扩张床吸附分离免疫球蛋白IgY

Separation and purification of immunoglobulin IgY with hydrophobic charge-induction expanded bed adsorption

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