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蓝色染料配基亲和纯化眼镜蛇心脏毒素的研究

Purification of Cardiotoxin of Naja naja atra with Cibacron Blue F3G-A Absorbents

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【作者】 刘浩然杨金菊戴平李荣秀

【Author】 LIU Hao-ran~1,YANG Jin-ju~2,DAI Ping~2,LI Rong-xiu~(1*)(1.Research Center of Biotechnology,Shanghai Institutes of Biology Sciences,Chinese Academy of Sciences,Shanghai 200233;2.East China Normal University School of Life Science,Shanghai 200062,P.R.China)

【机构】 中国科学院上海生命科学学院生物工程研究中心华东师范大学生命科学院中国科学院上海生命科学学院生物工程研究中心 上海200233上海200062上海200233

【摘要】 探索了蓝色染料 (CibacronBlueF3G -A)亲和分离中华眼镜蛇心脏毒素的可能性。采用环氧基活化法制备蓝色染料亲和介质 ,中性条件下提取眼镜蛇粗毒中的心脏毒素。Tricine系统SDS -PAGE多肽电泳和Lowry法蛋白定量分析纯化效果 ,发现蓝色染料琼脂糖一步纯化中华眼镜蛇心脏毒素的纯度达到 84 % ,结合量为 6 .9mg/ml介质。这是首次利用小分子亲和配基纯化心脏毒素。与生物大分子配基相比 ,活性染料分子具有价格便宜 ,易于合成 ,性质稳定 ,不易降解和适合大规模生产等优点。

【Abstract】 The purification of cardiotoxins with Cibacron Blue F3G-A Sepharose was developed.Blue sepharose was synthesized by the methods of epoxy-activate,distilled cardiotoxin from cobra venom on the condition of median pH.The samples were analyzed by peptide SDS-PAGE under tricine system and by protein concentration determined.It was shown that the one-step purification of cardiotoxin could get a purity of 84% with capacity of 6.9mg/g gel from Chinese cobra (Naja naja atra) crude venom.This is the 1st report of low molecular weight ligand for cardiotoxins purification other than immunoaffinity purification.Affinity adsorbents based on immobilized dyes offer important advantages circumventing many of the problems associated with biological ligands,which are resistant to both chemical and biochemical degradation and can be produced at an affordable price.

【基金】 “十五”国家重大科技专项 (2 0 0 2AA2Z345E)
  • 【分类号】Q599
  • 【下载频次】126
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