Preparation Of Immunomagnetic Beads And Its Preliminary Application

Immunomagnetic beads (Immunomagnetic beads,IMB) are uniform, polymer microspheres coated with ligand nanoscale materials. Immunomagnetic bead separation technology is based on the high uniformity of magnetic microspheres provide both a smooth hydrophobic surface to facilitate physical absorption of molecules, such as antibodies, antigens, etc. that allow immune binding of target materials. By the magnetic effect, specific target material are separated from the mixture. Immunomagnetic bead separation technology is a technology characterized by high efficiency, good repeatability, simple manipulation and cheap cost.In this study the chemical coprecipitation method is used for the synthesis of nano-Fe3O4 magnetic particles with a high degree of particle homogeneity, suspension stability, superparamagnetic, etc. The particle sizes were about 30～100 nm and the determination of the concentration of magnetic beads about 7.9 mg/mL, about size of about 30～100nm, wihich contribute to alkylating agent suspension and modification with aldehyde reagents enhance the performance of magnetic particles.Enterocolitica pestis is the pathogen of acute gastroenteritis-type food poisoning, foodborne pathogens are important, many countries have import and export of the bacteria as routine testing of food items. This study Yersinia enterocolitica recovery and expansion of antigen after culture preparation, through plate count to determine the concentration of about 2.3×1010 CFU/mL, this bacteria as the immunogen preparation of rabbit antiserum, using saturated crude ammonium sulfate method, over Sephadex G-200 purified IgG, coated with purified IgG immunomagnetic beads, IgG-coated beads to determine the optimal coupling conditions (including the best reaction medium, the best temperature, the best time). The establishment of enrichment using immunomagnetic beads technique Yersinia enterocolitica method to determine the optimal antibody concentration of coated beads, the best working concentration of immunomagnetic beads, immunomagnetic beads and the best role of bacteria time, the sensitivity of immunomagnetic bead technique, its sensitivity is about 70 CFU/mL. Antibody IgG-coated magnetic beads the best concentration of 100μg/mL, through the optimal reaction conditions was explored, and reduced immune bead enrichment Yersinia enterocolitica in the process of non-specific response. The results show that the use of immunomagnetic bead enrichment of bacteria in the process technology is an simple and effective technology. This study provided the basis for the construction of an rapid detection method to identify Yersinia enterocolitica.