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Praparation of Immunomagnefic Nanoparficles and Its Application in the Separation of CD34~+ Cell

Author: ZhaoLiNa
Tutor: ShenHeBai
School: Shanghai Normal University
Course: Physical and chemical
Keywords: Magnetic nanoparticles Nitrobenzaldehyde colorimetry Amino CD34 ~ cells CD34 antigen Hematopoietic stem / progenitor cells Umbilical Cord Blood
CLC: TB383.1
Type: Master's thesis
Year: 2008
Downloads: 141
Quote: 1
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Since nanotechnology generate nanotechnology has been widespread concern and rapid development which magnetic nanoparticles because of its uniform particle size, large specific surface area, the superparamagnetic advantages, favored by researchers in protein and immobilized enzyme immunoassays, RNA and DNA purification, cell separation and identification, targeting, play an increasingly important role in the administration and PCR. Hematopoietic stem cell transplantation to treat leukemia, lymphoma, multiple myeloma and other malignant diseases of the blood system, has an important significance in medicine. However a prerequisite step separating the high purity of the hematopoietic stem cells are hematopoietic stem cell transplantation, become the limiting factors for hematopoietic cell engineering. The hematopoietic stem cells have a variety of cell surface markers, but the identification of hematopoietic stem cells are the most commonly used is the CD34 antigen. CD34 monoclonal antibody may be the identification of specific hematopoietic stem cell surface CD34 antigen, therefore the use of the magnetic nanoparticles with CD34 monoclonal antibody binding forming a magnetic nanoparticle - antibody complexes the immune magnetic nanoparticles, the rapid separation of a high applied magnetic field The purity of the CD34 cells. This paper studies the immune Magnetic Nanoparticles and CD34 cells were isolated from umbilical cord blood. (1) with superparamagnetic γ-Fe2O3 nanoparticles synthesized by co-precipitation method, by TritonX-100, n-hexanol, cyclohexane-posed inverse microemulsion system, the TEOS ammonia catalytic The hydrolysis of the surface of the γ-Fe2O3 wrapped layer having a biological affinity for the silica layer to form a core-shell type magnetic nanoparticles. Having active amino functional group on its surface modified with aminosilane. TEM showed that the magnetic nanoparticles was a uniform particle size and good monodispersibility; the Infrared Characterization amino has been successfully modified at the surface of the magnetic nanoparticles. Aminated magnetic nanoparticles by a chemical reaction with antibodies, the formation of magnetic nanoparticle - antibody complexes, and through the UV characterization. Magnetic nanoparticles with different antibodies connection can be separated from the different cells, and provide a means for the separation of biological cells. (2), nitrobenzaldehyde colorimetric assay determination of amino quick and easy, and non-toxic, has a lot of advantages compared with other methods. Adoption of nitrobenzaldehyde nanometer magnetic particle interface amino quantitative colorimetric assay of the subsequent quantitative connection biological molecules is very important; nitrobenzaldehyde colorimetric assay also help to study the magnetic nanoparticles under different reaction conditions The amount of surface interface amino reactive silane interface helps to optimize and need further study the influence of the reaction conditions in the production of high-performance materials. (3) magnetic nanoparticles-CD34 antibody complex was added to the need to separate the samples, magnetic nanoparticles can be carried out with the target cell-specific binding, and then the external magnetic field on the attached the magnetic nanoparticles target cell retention. thereby achieve separation. The results showed the use of magnetic nanoparticles - antibody complex can be quickly and efficiently CD34 ~ cells from umbilical cord blood the grab elected separated CD34 ~ cells remain normal morphology, a high proliferative capacity and differentiation ability, indicating magnetic nanoparticle - antibody complex was separated CD34 ~ cell technology has a good effect.

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