摘要
目的 :分离除去蚓激酶白蛋白纳米粒溶液中未成粒的BSA ,利于FITC标记在纳米粒上 ;分离除去未与蚓激酶纳米粒结合的FITC ,降低FITC对检测荧光纳米粒的影响。方法 :采用SephadexG - 1 0 0和SephadexG - 5 0凝胶柱分离纯化。结果 :流速为 3ml/min时 ,SephadexG - 1 0 0柱 (2 1 .5cm ,φ1 .9cm)能够分离除去 (2 9.3± 3.6 ) %未形成纳米粒的BSA ;流速为 0 .2 5ml/min时 ,SephadexG - 5 0凝胶柱 (1 0cm ,φ2cm)可以完全分离除去未与纳米粒结合的FITC。结论 :通过分离 ,FITC有效地标记在纳米粒上 ;降低了背景荧光 ,使荧光纳米粒在荧光显微镜下清晰显现。
Objective: To separate the unencapsuled BSA from albumin nanoparticles of lumbrokinase in the solution so as to ensure FITC labelled on the nanoparticles better; to remove the free FITC from labeling process and to reduce FITC's affection on the detection of nanoparticles. Methods: The constituents were isolated on Sephadex G-100 and Sephadex G-50 gel column chromatography. Results: The (29.3±3.6) % unencapsuled BSA in the solution was separated and removed by using Sephadex G-100 gel column (21.5cm, φ 1.9cm)while flow rate was 3ml/10min. FITC was removed completely by using Sephadex G-50 gel column (10cm, φ 2cm) While flow rate was 0.25ml/10min. Conclusion: Through separation, FITC can be effectively labeled on nanoparticles; the background fluorescence was reduced, and the nanoparticles labeled with FITC can be detected easily under fluorescence-microscope.
