纳米免疫磁性微球的制备和性能研究

Preparation of immunomagnetic beads and study of its function

目的:初步探讨纳米免疫磁性微球(Immunomagneticbeads,IMB)的制备方法及其性能。方法:用FeC纳米磁粉为内核,用反相乳液法制备壳聚糖磁性复合微球,用戊二醛活化复合微球并交联鼠抗人CD3单克隆抗体,制备出人CD3纳米免疫磁性微球,用人CD3纳米磁性免疫微球富集人外周血中CD3+细胞,并检测富集细胞的效果。结果:壳聚糖磁性复合微球平均粒径为560nm,粒径分布在409～710nm之间,用此微球制备的人CD3免疫磁性微球蛋白联接率达93.8%,流式细胞术检测人CD3纳米免疫磁性微球分离CD3+细胞纯度达到94.8%。结论:制备的人CD3纳米免疫磁性微球具有分离细胞纯度高,细胞损伤小,不用解离磁球即可检测等特点。

Objective：To explore preliminarily the method of the preparetion of the immunomagnetic beads（IMB） by Fe/C nanomagnefic particles covered with chitosan and coupled with CD3 monoclone antibody. Methods：Chitosan magnetic composite microspheres were prepared by reverse microemulsion. The composite microspheres were activated by glutaraldehyde and coupled with human CD3 antibody. Then the self-prepared IBM were acquired. CD3 cells were separated from human blood by self-prepared immunomagnefic beads. And the effect of the separation of CD3 cell was detected. Results： The average diameter of self-prepared chitosan magnetic composite microspheres was 560 nm, the diameter distributing of composite microsphere was from 409 nm to 710 nm. With self-prepared IMB, the purity of 95.8% of CD3 cells was harrested. Conclusion：With self-prepared CD3 immunomagnetic bead（IMB）, high 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 IMB.