凝胶层析和离子交换层析结合法纯化重组降血压肽VLPVPR
Purification of recombinant antihypertensive peptides VLPVPR by gel and ion exchange chromatography
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摘要: 为了提高VLPVPR的纯度,采用凝胶层析结合离子交换层析的方法对VLPVPR进行纯化。先用Sephadex G-10凝胶对VLPVPR溶液脱盐,再考察离子交换剂(SP Sepharose Fast Flow和Q Sepharose Fast Flow)对重组降血压肽VLPVPR的纯化效果,确立纯化VLPVPR的优化工艺。实验结果表明SP Sepharose Fast Flow不适于VLPVPR的纯化。采用Q Sepharose Fast Flow的纯化工艺为:上样量为柱床体积的10%,用10mmol/L pH9.0的CHES缓冲液洗脱,流速为1mL/min。VLPVPR回收率为93.8%,纯度达到47.2%。采用Q Sepharose Fast Flow纯化提高了VLPVPR的纯度。Abstract: To purify the peptide VLPVPR, gel filtration combined with ion exchange chromatography method was employed.Sephadex G-10 was used to desalt firstly.Compared the purification effective of SP Sepharose Fast Flow and Q Sepharose Fast Flow.SP Sepharose Fast Flow was not suitable for VLPVPR purification while Q Sepharose Fast Flow was suitable for VLPVPR purification.The optimization condition of Q Sepharose Fast Flow were pH9.0, feeding volume 10% BV, CHES concertration 10 mmol /L, elute rate 1.0mL /min.The recovery ratio was 93.8% and the purity ratio was 47.2%.The purification effective of Q Sepharose Fast Flow was improved.