汽巴蓝F3GA染料亲和色谱介质的制备及其对黑芸豆凝集素的特异性吸附

doi:10.7506/spkx1002-6630-201823008

食品科学 ›› 2018, Vol. 39 ›› Issue (23): 47-53.doi: 10.7506/spkx1002-6630-201823008

汽巴蓝F3GA染料亲和色谱介质的制备及其对黑芸豆凝集素的特异性吸附

赵金龙1，李延红1，孙汉巨1，方利2，孙先保1，唐明明1，张扬3，武香玉4，何述栋1,*

1.合肥工业大学食品与生物工程学院，安徽合肥 230009；2.常州天地人和生物科技有限公司，江苏常州 213000；3.河南省农业科学院经济作物研究所，河南郑州 450002；4.山东省肥城市食品药品监督管理局，山东肥城 271608

出版日期:2018-12-15 发布日期:2018-12-17
基金资助:
国家自然科学基金青年科学基金项目（31701524）；国家自然科学基金面上项目（31771974）；安徽省自然科学基金面上项目（1708085MC70）；合肥工业大学中央高校专项基金项目（JZ2018HGTB0245）；合肥工业大学实验室自制仪器设备项目（Z201619）

Preparation of Cibacron Blue F3GA Affinity Chromatography Medium and Its Specific Adsorption toward Lectin from Black Kidney Beans

ZHAO Jinlong1, LI Yanhong1, SUN Hanju1, FANG Li2, SUN Xianbao1, TANG Mingming1, ZHANG Yang3, WU Xiangyu4, HE Shudong1,*

1. School of Food and Biological Engineering, Hefei University of Technology, Hefei 230009, China; 2. Smart-Lifesciences Co. Ltd., Changzhou 213000, China; 3. Industrial Crops Research Institute, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China; 4. Feicheng Food and Drug Administration, Feicheng 271608, China

Online:2018-12-15 Published:2018-12-17

摘要/Abstract

摘要： 采用反相悬浮再生法制备6 g/100 mL的琼脂糖微球，采用环氧氯丙烷对琼脂糖微球进行二次交联，并进行染料配体汽巴蓝F3GA的偶联，得到一种新型活性Blue Beads 6FF微球。通过傅里叶变换红外光谱、扫描电子显微镜、压力流速曲线、溶胀率、泄漏率和孔隙率等对微球物理性能进行表征。结果发现，微球球形度较好，粒径大小均一，平均粒径为56.37 μm左右，具有较高的孔隙率、溶胀率及广范围的pH值稳定性，且耐压能力强。以黑芸豆凝集素为目标蛋白，通过Langmuir吸附等温式拟合，结果表明该填料对黑芸豆蛋白有较高的吸附量（19.7 mg/mL），微球吸附性能良好，可实现对凝集素蛋白的快速分离和纯化。

关键词: 琼脂糖微球, 反相悬浮再生法, 汽巴蓝F3GA, 亲和色谱, 黑芸豆凝集素

Abstract: Agarose microspheres (6 g/100 mL) were prepared by reversed-phase suspension regeneration method and they were cross-linked twice with epichlorohydrin and then coupled with the dye Cibacron Blue F3GA as a special ligand to form novel active microspheres (Blue Beads 6FF). The properties of the dye affinity medium were characterized by Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), pressure against flow velocity curve, swelling rate, leakage rate, and porosity. Our results showed that the microspheres had a good sphericity and uniform size with high porosity and swelling rate, a wide range of pH stability and strong pressure resistance. The average particle size of the microspheres was approximately 56.37 μm. The adsorption properties of the dye affinity medium were evaluated for lectin purification from black kidney beans. By fitting the typical Langmuir adsorption isotherm, the equilibrium adsorption capacity of lectin from black kidney beans was determined as 19.7 mg/mL microspheres. The dye affinity microspheres enabled rapid isolation and purification of lectin.

Key words: agarose microspheres, reversed-phase levitation regeneration method, Cibacron Blue F3GA, affinity chromatography, lectin from black kidney beans

中图分类号:

Q652.6

赵金龙，李延红，孙汉巨，方利，孙先保，唐明明，张扬，武香玉，何述栋. 汽巴蓝F3GA染料亲和色谱介质的制备及其对黑芸豆凝集素的特异性吸附[J]. 食品科学, 2018, 39(23): 47-53.

ZHAO Jinlong, LI Yanhong, SUN Hanju, FANG Li, SUN Xianbao, TANG Mingming, ZHANG Yang, WU Xiangyu, HE Shudong. Preparation of Cibacron Blue F3GA Affinity Chromatography Medium and Its Specific Adsorption toward Lectin from Black Kidney Beans[J]. FOOD SCIENCE, 2018, 39(23): 47-53.

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汽巴蓝F3GA染料亲和色谱介质的制备及其对黑芸豆凝集素的特异性吸附

Preparation of Cibacron Blue F3GA Affinity Chromatography Medium and Its Specific Adsorption toward Lectin from Black Kidney Beans

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