抗sHLA-Ⅰ免疫磁珠制备及鉴定.doc

抗sHLA-Ⅰ免疫磁珠制备及鉴定[摘要] 目的 制备抗sHLA-Ⅰ免疫磁珠去除红细胞悬液和机采血小板中的sHLA-Ⅰ分子。方法 将单克隆抗体W6/32以共价偶联的方式包被制备免疫磁珠，以FITC标记的羊抗鼠二抗标记磁珠，用流式细胞仪鉴定包被效果。取10例红细胞悬液和机采血小板上清和制备好的免疫磁珠孵育后移去磁珠，用ELISA检测吸附前后sHLA-Ⅰ浓度变化评价吸附效果。结果 制备的抗sHLA-Ⅰ免疫磁珠包被效率约为75%。对红细胞悬液和机采血小板中sHLA-Ⅰ的吸附效果可达到84%和83%。 结论 采用单克隆抗体W6/32制备的免疫磁珠能有效去除红细胞悬液和机采血小板中的sHLA-Ⅰ分子，在改善临床输血安全方面有广阔的应用前景。 [关键词]可溶性组织相容性抗原Ⅰ类分子；免疫磁珠 [中图分类号] R392.33 [文献标识码]B[文章编号] 1673-9701（2011）19-149-02 The Preparation and Appreciation of Anti-sHLA-I Immune Magnetic Bead YAN Shaorong1 LI Qiang1 YU Shifang2 1. Department of Clinical Laboratory，Ruian City Peoples Hospital，Ruian 325200，China；2.Blood Bank，First Affiliated Hospital of Wenzhou Medical College，Wenzhou 325000，China [Abstract] Objective To prepare anti-soluble human leukocyte antigens I （sHLA-I） immune magnetic bead for adsorbing and removing the soluble HLA ?I of red cell suspension and machine adopting platelet. MethodsThe magnetic beads are coated with monoclonal antibody W6/32 by covalence coupling’s way. Then the magnetic beads are marked by the sheep-anti-mouse second antibody marked by FITC and are appraised the coating effect by flow cytometer（FCM）. Ten example red cell suspension and machine adopting platelet supernatant are selected and incubated with the prepared immune magnetic bead. Then the beads are removed from the mixed liquor and the density of the sHLA-I in the supernatant is detected by ELISA to compare the density change before and after adsorbing and appreciate the adsorbing effects. Results The prepared anti-sHLA-I magnetic beads?coating efficiency is 75%.The adsorbing effect to soluble HLA-I existing in red cell suspension and machine adopting platelet respectively is 84% and 83%. Conclusion The magnetic beads prepared with monoclonal antibody W6/32 can adsorb and remove efficiently the soluble HLA-I existing in red cell suspension and machine adopting platelet，which will have extensively appliedperspective in improving clinical transfusing blood safe. [Key words] Solubl