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亲和层析法从红芸豆中纯化红血球凝集素的研究
引用本文:廖涛,张芹,蒲铃铃. 亲和层析法从红芸豆中纯化红血球凝集素的研究[J]. 成都医学院学报, 2010, 5(2): 145-148. DOI: 10.3969/j.issn.1674-2257.2010.02.015
作者姓名:廖涛  张芹  蒲铃铃
作者单位:成都医学院生物医学系生物技术专业,四川,成都,610083;成都医学院生物医学系基础医学院实验教学中心,生物技术实验室,四川,成都,610083
基金项目:成都医学院实验室开放基金
摘    要:目的制备一种亲和树脂快速分离红血球凝集素。方法通过甲状腺球蛋白和树脂的耦联来从红芸豆粗提液中快速分离红血球凝集素。结果从红芸豆粗提液中快速地提取出了较纯的红血球凝集素,经SDS-PAGE上显示亚基分子量为32kD.并通过质谱确认为红血球凝集素,提取得到的凝集素使人红细胞50%凝集的蛋白质最低浓度为4μg/ml左右。结论成功制备亲和树脂并用于快速分离红血球凝集素。

关 键 词:凝集素  亲和树脂  红芸豆  分离纯化

Purification of erythrohemagglutinin from Phaseolus vulgaris using affinity resin
LIAO Tao,ZHANG Qin,PU Ling-ling. Purification of erythrohemagglutinin from Phaseolus vulgaris using affinity resin[J]. Journal of Chengdu Medical College, 2010, 5(2): 145-148. DOI: 10.3969/j.issn.1674-2257.2010.02.015
Authors:LIAO Tao  ZHANG Qin  PU Ling-ling
Affiliation:1.Department of Biotechnology,Chengdu Medical College,2.Experimented Teaching Center of Basic Medical Science Department of Biotechnology laboratory,Chengdu Medical College,Chengdu,Sichuan 610083,China)
Abstract:Objective To study a new method for purifying erythrohemagglutinin (PHA-E) production from Phaseolus vulgaris by affinity resin.Methods Affinity resin was made by Sepharose CL 4B connecting with thyroglobulin,and it was used to purify PHA-E from the excellent seeds of Phaseolus vulgaris. The final gel filtration chromatography,purity,molecular weight and isoelectric point of purified PHA-E were determined by electrophoresis and mass spectrum.Results The purified PHA-E showed single PAGE outline and had apparent subunit molecular weights of 32 kD by SDS-polyacrylamide gel electrophoresis(SDS-PAGE). PHA-E could promote agglutination of human erythrocytes evenat 4 μg/ml.Conclusion A rapid method for purifying PHA-E production from Phaseolus vulgaris by affinity resin is established successfully.
Keywords:erythrohemagglutinin  affinity resin  Phaseolus vulgaris  separation and purification
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