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| Chemical Modification of Lysostaphin with Activated Polyethlene Glycol |
| WANG Yong1, LIU Mu-rong2, WAN Hai-tong1, LIU Xiang2, ZHANG Jia-hui2, FENG Xiao-yan2, ZHANG Fan1 |
1. Biological Engineering Institute of Zhejiang Chinese Medical University, Hangzhou 310053, China;
2. BIODOOR Biotechnology Co., Ltd of Hangzhou City in Zhejiang Province, Hangzhou 310011, China |
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Abstract Objective: To study the reaction conditions for the chemical modification of recombinant lysostaphin (Lysostaphin) with activated polyethlene glycol (PEG) and the methods for the purification of PEG-Lysostaphin. Methods: Disrupt the cell wall of bacteria by ultrasonic cell disruptor and purify the lysostaphin by Cation exchange chromatography (SP) and Hydrophobic chromatography (HIC); Lysostaphin was chemical modified with activated mono-methoxy polyethlene glycol succinimide propionate (mPEG- SPA) under various conditions, the ration of mono-mPEG-Lysostaphin was determined by SDS-PAGE and EMALDI-TOF-MS; The reaction mixture was purified by Sephacryl S-200 molecular size exclusion chromatography. Results: The optimal reaction was pH 8.0,temperature 4℃, 1:5 for the mass ratio of lysostaphin and mPEG, two hours for reaction time was selected. The reaction mixture was preliminarily separated by one step Sephacryl S-200 molecular size exclusion chromatography. Conclusion: It’s preliminarily determined that the optimal reaction conditions for the chemical modification of lysostaphin with activated polyethlene glycol and the methods for the purification of PEG-Lysostaphin.
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Received: 29 January 2013
Published: 25 June 2013
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