生物学杂志

生物学杂志

• 技术方法 • 上一篇    下一篇

人脐带血血红蛋白的纯化

  

  1. 天津工业大学 环境与化学工程学院， 天津 300387
  • 出版日期:2016-08-18 发布日期:2016-08-18
  • 通讯作者: 王 翔，副教授，研究生导师，主要研究方向为聚合血红蛋白方面的研究，E-mail：tpuwx@163.com
  • 作者简介:陈晨，研究生，主要研究方向为血红蛋白的纯化，E-mail:cchentpu@yeah.net
  • 基金资助:
    国家青年科学基金（No 21202117/B020703）

The purification of placenta hemoglobin

  1. School of Environment and Chemical Engineering, Tianjin Polytechnic University, Tianjin 300387, China
  • Online:2016-08-18 Published:2016-08-18

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摘要： 为了探索人脐带血血红蛋白的纯化方法，用0.9%生理盐水洗涤脐带血，经蒸馏水进行粗提，经Q300阴离子交换纯化血红蛋白，对纯化后的血红蛋白进行浓缩，得到终产品。测定血红蛋白浓度及高铁血红蛋白含量，并用SDS-PAGE和HPLC进行纯度分析，通过血样分析仪进行氧饱和曲线分析。纯化和浓缩后血红蛋白浓度为11.5 g/dl，高铁血红蛋白含量≤1%；经SDS-PAGE和HPLC分析血红蛋白纯度较高，不含杂蛋白，凝胶成像系统分析纯度为99.10%±0.70%；纯化前后血红蛋白的氧合曲线无明显偏移，不影响其载氧活性。研究证明，成功地得到了人脐带血血红蛋白纯化的方法。

关键词: 人脐带血, 离子交换, 纯化

Abstract: The aim of the study is to establish the purification methods of placenta hemoglobin. The placenta hemoglobin was eluted by 0.9% Na chloride(NS) and extracted by distilled water, which was further purified by Q300 anion-exchange chromatography, and then the final products was got by concentrating the purified hemoglobin. The concentration of hemoglobin and the content of methemoglobin of the final products was measured, and the purity was analyzed by SDS-PAGE and HPLC,the oxygen saturation curve was analyzed through the blood analyzer.The concentration of final products was 11.5 g/dl,and the content of methemoglobin is 1% or less; hemoglobin had a higher purity by the analysis of SDS-PAGE and HPLC, excluding mixed protein, the purity was 99.10%±0.70% analyzed by Gel Imaging System; the oxygenation of hemoglobin curve had no significant deviation after purification, the activity of carrying oxygen would not be affected. The research shows that the methods to purify placenta hemoglobin were established.

Key words: placenta hemoglobin, ion exchange, purification