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Preparation and application of streptavidin magnetic particles
作者单位:ZHANG ZhiFeng(Biochip Research and Development Center, Northwest University (National Engineering Research Center for Miniaturized Detection System), Xi'an 710069, China) ; ZHU HongLi(Biochip Research and Development Center, Northwest University (National Engineering Research Center for Miniaturized Detection System), Xi'an 710069, China) ; TANG YiTong(Biochip Research and Development Center, Northwest University (National Engineering Research Center for Miniaturized Detection System), Xi'an 710069, China) ; CUI Ting(Shaanxi Lifegen Co., Ltd., Xi'an 710069, China) ; GENG TingTing(Shaanxi Lifegen Co., Ltd., Xi'an 710069, China) ; CHEN Chao(Biochip Research and Development Center, Northwest University (National Engineering Research Center for Miniaturized Detection System), Xi'an 710069, China;Shaanxi Lifegen Co., Ltd., Xi'an 710069, China) ; CUI YaLi(Biochip Research and Development Center, Northwest University (National Engineering Research Center for Miniaturized Detection System), Xi'an 710069, China) ;
基金项目:国家自然科学基金;国家高技术研究发展计划(863计划)
摘    要:Two kinds of streptavidin magnetic particles,namely streptavidin GoldMag particles and streptavidin amino terminal particles were prepared by the methods of physical adsorption and covalent interaction respectively.The streptavidin coated on magnetic particle surface,crucial to many applications,was greatly influenced by the choice of the different buffer.Compared with DynalbeadsM-270 streptavidin, the binding capacity for biotin of different streptavidin magnetic particles was determined by enzyme inhibition method,and the coupling capacity and activity of biotinylated oligonucleotide on their sur- face were also analyzed.The results indicated that the streptavidin GoldMag particle prepared by physical adsorption was stable in STE(NaCl-Tris-EDTA)buffer that was frequently used in nucleic acid hybridization and detection.The streptavidin amino terminal particles prepared by covalent interaction could be used both in STE buffer and PBS(phosphate buffered saline)buffer.The biotin binding ca- pacity for 1 mg of streptavidin GoldMag particles and streptavidin amino terminal particles was 4950 and 5115 pmol respectively.The capacity of biotinylated oligonucleotide(24 bp)coupled on 1 mg of GoldMag and amino terminal magnetic particles was 2839 and 2978 pmol separately.These data were about 6-7 times higher than those of DynabeadsM-270 streptavidin.The hybridization results with FITC-labeled complementary probe on magnetic particle surface demonstrated that the oligonucleotide coupled on streptavidin magnetic particles had high biological activity.

收稿时间:15 October 2005
修稿时间:30 March 2006

Preparation and application of streptavidin magnetic particles
Authors:Zhang ZhiFeng  Zhu HongLi  Tang YiTong  Cui Ting  Geng TingTing  Chen Chao  Cui YaLi
Institution:1. Biochip Research and Development Center, Northwest University (National Engineering Research Center for Miniaturized Detection System), Xi'an 710069, China
2. Shaanxi Lifegen Co., Ltd., Xi'an 710069, China
3. Biochip Research and Development Center, Northwest University (National Engineering Research Center for Miniaturized Detection System), Xi'an 710069, China;Shaanxi Lifegen Co., Ltd., Xi'an 710069, China
Abstract:Two kinds of streptavidin magnetic particles, namely streptavidin GoldMag particles and streptavidin amino terminal particles were prepared by the methods of physical adsorption and covalent interaction respectively. The streptavidin coated on magnetic particle surface, crucial to many applications, was greatly influenced by the choice of the different buffer. Compared with Dynalbeads®M-270 streptavidin, the binding capacity for biotin of different streptavidin magnetic particles was determined by enzyme inhibition method, and the coupling capacity and activity of biotinylated oligonucleotide on their surface were also analyzed. The results indicated that the streptavidin GoldMag particle prepared by physical adsorption was stable in STE (NaCl-Tris-EDTA) buffer that was frequently used in nucleic acid hybridization and detection. The streptavidin amino terminal particles prepared by covalent interaction could be used both in STE buffer and PBS (phosphate buffered saline) buffer. The biotin binding capacity for 1 mg of streptavidin GoldMag particles and streptavidin amino terminal particles was 4950 and 5115 pmol respectively. The capacity of biotinylated oligonucleotide (24 bp) coupled on 1 mg of GoldMag and amino terminal magnetic particles was 2839 and 2978 pmol separately. These data were about 6–7 times higher than those of Dynabeads®M-270 streptavidin. The hybridization results with FITC-labeled complementary probe on magnetic particle surface demonstrated that the oligonucleotide coupled on streptavidin magnetic particles had high biological activity.
Keywords:streptavidin  magnetic particles  preparation  application
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