色谱

色谱

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固定化铜离子亲和膜色谱柱吸附血红蛋白的研究

秦晓蓉,伍林,易德莲,胡雷,曹淑超   

  1. Applied Chemistry Research Institute, Wuhan University of Science and Technology, Wuhan 430081, China
  • 收稿日期:2004-07-09 修回日期:1900-01-01 出版日期:2005-05-30 发布日期:1988-09-25
  • 通讯作者: 秦晓蓉
  • 基金资助:

Study of Adsorption for Hemoglobin by Immobilized Copper Ion Affinity Membrane Chromatographic Column

QIN Xiaorong, WU Lin, YI Delian, HU Lei, CAO Shuchao   

  1. Applied Chemistry Research Institute, Wuhan University of Science and Technology, Wuhan 430081, China
  • Received:2004-07-09 Revised:1900-01-01 Online:2005-05-30 Published:1988-09-25

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摘要： 将纤维素滤纸进行碱处理及环氧活化、偶联亚氨基二乙酸、固定化铜离子等处理，并将其装入自制的色谱柱管,制得固定化铜离子亲和膜色谱柱。该柱可用于吸附血红蛋白(hemoglobin,Hb),吸附率可达到90%以上。考察了上样量、pH值、温度、上样速度等因素对固定化铜离子亲和膜吸附Hb的影响。实验结果表明,固定化铜离子亲和膜色谱柱吸附血红蛋白的最佳条件为：室温下实验,缓冲体系的pH值控制在6~8,上样速度0.5~1.0 mL/min，上样量为3.16~7.90 mg/g。

关键词: 固定化铜离子, 亲和层析, 亲和膜, 吸附 , 血红蛋白

Abstract: Immobilized copper ion affinity membrane was prepared by using cellulose filter paper as matrix. The membrane was loaded into a chromatographic column designed in the laboratory and the immobilized copper ion affinity membrane chromatographic column was prepared. Hemoglobin (Hb) was adsorbed by the column and its adsorption rate was above 90%. The experimental process was as follows: a set of 5 immobilized copper ion affinity membranes were loaded into a column, uncombined copper ion was removed with 5 mL of sodium phosphate-citric acid buffer (pH 7); the Hb sample volume was 2 mL, and uncombined Hb was eluted with 10 mL of the buffer. The Hb concentration was determined by ultraviolet spectrophotometry at 405 nm. The major influence factors on hemoglobin adsorption rate such as Hb sample amount (1.58-31.6 mg/g (2 mL of sample volume)), pH value range (3.6-10.6), temperature range (25-50 ℃), and sample flow-rate (0.5-2.5 mL/min) were investigated. The optimal operation conditions were found as follows: the Hb sample amount of 3.16-7.90 mg/g, sample flow-rate of 0.5-1.0 mL/min, test temperature of room temperature, pH value range of 6-8. Compared with aqueous two-phase system, liquid-solid extraction system and the conventional column method, the membrane medium is cheaper and easier to be scaled up.

Key words: adsorption , affinity chromatography, hemoglobin, immobilized copper ion, affinity membrane