Preparation And Characterization Of Chitosan Magnetic Composite Microspheres And Its Application In Enrichment Of T Cells

The immunomagnetic microspheres (IMM) were prepared by covering the magnetic composite microspheres with antibody or biomolecule by physical adsorption or covalent crossing. Immunomagnetic microspheres could link specially to a kind of biomolecule or the antigen on the surface of cells, as passing throught magnetic field , the cells and biomolecule which combined with immunomagnetic microspheres is stayed in magnetic field, then the target cells or biomolecule could be separated from other kind of cells or biomolecule. This process is called immunomagnetic separation. Immunomagnetic separation quickly spread to the fields of physiology, pathology, pharmacology, microbiology, et al for the advantage of convenience, quick, innocuity (compare with traditional methods). The following progress were acquired in the text begin with the preparation of chitosan magnetic composite microspheres.1. By reverse microemulsion, the chitosan magnetic composite microspheres (CMCM) were prepared and characterized in appearance . magnetic response, content of Fe. amount of groups . and so on. Also polystyrene magnetic composite microspheres (PMCM) were prepared and characterized and compared with chitosan microspheres. It was showed that CMCM were superior carrier for immunomagnetic microspheres.2. According to the experiments, some factors (temperature, time, the ion strength, the condition of activation) were considered in the course of protein adsorption of CMCM. It was found that the temperature and time were the subordinate factors for behavior of adsorption, the other two factors (ion strength , the condition of activation)were critical factors to it. The amount of adsorption of Albumin Bovine(BSA) enhanced nearly 5 times from 5.4mg/g to 24.6mg/g. The adsorption had reverse ratio with concentration of NaCl. The situation that activated CMCM in low ion strenghth was commended in the experiments.3. After activated by glutaraldehyde, the CMCM were coupled with the CD₃ antibody, The CD₃ immunomagnetic microspheres were prepared. CD₃⁺ cells were separated from human blood by self-prepared immunomagnetic microspheres. And theeffect of the separation of CD3+ cells were detected by Flow CytoMeter (FCM) . With self-prepared CD3 immunomagnetie microspheres, 95% purity of CD3+ cells was obtained. Further more the CD3+ cells have hardly any damnify, and could be detected through FCM without cutting off the 1MB.