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Preparation of Magnetic Monodisperse Microspheres for Plasmid DNA Purification

Author: ZhangHaiPing
Tutor: BaiZuo
School: Tianjin University
Course: Biochemical Engineering
Keywords: Magnetic Polymer Microspheres Dispersion polymerization Permeation method Plasmid Purification Free emulsion polymerization method Swelling method
CLC: Q75
Type: Master's thesis
Year: 2007
Downloads: 179
Quote: 0
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Abstract


Bioactive substances using magnetic polymer microspheres separation technology , is a highly efficient integrated technology, this technology is the future direction of bioseparation technology development . This article aims to prepare a high degree of crosslinking of superparamagnetic monodisperse magnetic polymer microspheres , and applied to the separation and purification of biological macromolecules , such as proteins , plasmid . Article of glycidyl methacrylate (GMA) as the monomer and divinylbenzene (DVB) as a crosslinking agent, using dispersion polymerization combined permeate prepared by the degree of crosslinking is 3-5% , the particle size of 1-1.5 rigid μm monodisperse polymer microspheres , and the successful introduction of the amino group of the high-density ; characterize the corresponding properties of the magnetic polymer microspheres with PSA, ESEM, VSM, IR analytical tools , and the results show the preparation of micro pellets diameter uniformity , morphological integrity , non-porous surface is smooth , with superparamagnetic more than the the saturation magnetization 16.48emu / g , amino-modified density 4.432mmol / g dry resin . In order to test the effect of the practical application of the microspheres , the use of the preparation of magnetic microspheres of plasmid DNA were isolated and purified. Microspheres fast adsorption process , 1min can reach equilibrium ; in the pH 6.5, the salt concentration of 0.5mol / L under the adsorption conditions , the maximum amount of adsorption can be achieved 90.35μg/mg Dry RESIN ; at pH 9, the salt concentration of 1.25mol / L the elution volume 400μL conditions the elution of nucleic acid recovery reached 97.2%; microspheres reuse exhibit very high stability in operation , but also having a good chemical stability . Microspheres purified plasmid DNA . The application directly on alkaline lysis of the cells in liquid adsorption , 1.5 ml broth in the separately recovered 19.4μg 24.8μg ( by adding RNA enzymes ) plasmid . The results show that the magnetic microspheres can be quick and easy , purified plasmid DNA . In order to obtain a smaller particle size , the paper has a soap-free emulsion polymerization method, combined with swelling method successfully synthesized 2-5.5 % degree of crosslinking of 200-600nm narrow - dispersed microspheres ratio the the saturation magnetization 10.99emu / g .

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CLC: > Biological Sciences > Molecular Biology > Molecular Genetics
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