Purification of Phytase from Transgenic Maize by Immunoaffinity Chromatography

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China Biotechnology ›› 2013, Vol. 33 ›› Issue (6) : 125-130.

ZHAO Qian-qian¹, ZHOU Xiao-jin¹, LIU Xiao-qing¹, YANG Wen-zhu¹, LI Su-zhen², LIU Qi³, CHEN Ru-mei¹

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Abstract

For further investigating the enzymatic characterizations, evaluating the allergenicity and feed safety of the phytase generated from transgenic phytase corn, A more effective purification system using immuno-affinity chromatography was established. In order to increase the loading capacity and maintain the specificity, four monoclonal antibodies recognizing different epitopes of phytase were combined together. The crude extraction could be concentrated by 80% ammonium sulfate, and then subjected to dialysis and immune-affinity chromatography. The purification results showed that the obtained protein has an exclusive specific band in SDS-PAGE and 470.99 U/mg in specific activity. Furthermore, comparing to ion exchange chromatography, the immune-affinity based method was more stable and effective, the resultant product can meet the demands for various analysis.

Key words

Phytase / Protein purification / Immune-affinity chromatography / Sulfate precipitation / Ion exchange chromatography

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ZHAO Qian-qian, ZHOU Xiao-jin, LIU Xiao-qing, YANG Wen-zhu, LI Su-zhen, LIU Qi, CHEN Ru-mei. Purification of Phytase from Transgenic Maize by Immunoaffinity Chromatography[J]. China Biotechnology, 2013, 33(6): 125-130

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