Preparation and application of streptavidin magnetic particles

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.