摘要
目的:建立125Ⅰ标记抗巨噬细胞移动抑制因子(MIF)单克隆抗体(mAb)方法,探讨其在体内外生物学活性。方法:①利用Iodogen法,Na125Ⅰ标记Anti-mMIF mAb,用Sephadex G-25柱凝胶过滤层析法分离纯化,测定其标记率。纸层析法测定放射化学纯度和稳定性。②采用ELISA和改良MMI鉴定其免疫活性及生物学活性。③观察小鼠体内的生物学分布情况。结果:①标记的最佳条件为Iodogen 40~100μg、抗体20~50μg[Iodogen(m)∶抗体(m)=2∶1],加入Na125I15~30 MBq、室温反应10~15 min,标记率为(90.40±3.34)%,放射化学纯度为(97.60±1.14)%,比活度为12.2~26.0 MBq/μg;标记物4℃条件下放置3周后放射化学纯度仍为90.36%。②125Ⅰ-mMIF mAb能与抗原MIF特异性结合,较好地保持其免疫活性及生物学活性。③体内生物学分布,在肺、肝、脾、肾内具有较高的放射性计数。结论:Iodogen法获得高标记率和放射化学纯度的125Ⅰ-mMIF mAb,具有很好的体外稳定性,其在体内外保持较好的生物学活性。为进一步应用125I-mMIF mAb进行放射免疫显像和靶向治疗奠定了实验基础。
AIM： Find the method to make 125Ⅰ-labeled anti-Macrophage migration inhibitory factor（MIF） mAb and investigate its in vivo biologic activity.METHODS： ①Anti-MIF mAb was radioiodinated with Na125Ⅰ by Iodogen method followed by purified using Sephadex G-25 gelfiltration chromatography.The labeling efficiency of 125Ⅰ-labeled anti-MIF mAb was measured.The radiochemicalpurity and the stability were assayed by paper chromatograph.②The immunocompetence and the in vivo biologic activity were evaluated by using ELISA and modified MMI.③The in vivo biodistribution of 125Ⅰ-labeled anti-MIF mAb was detected using mouse model.RESULTS： ①The best reaction condition for labeling was： Mixed 40-100 μg Iodogen with 20-50 μg anti-MIF mAb（Iodogen（m）： anti-MIF mAb（m）=2∶1）;added 15-30 MBq Na125Ⅰ to Iodogen-Ab mix;kept reaction at RT for 10-15 min.The labeling efficiency of 125Ⅰ-labeled anti-MIF mAb was（90.40±3.34）%.The radiochemicalpurity was（97.60±1.14）%.And the specific activity was 12.2-26.0 MBq/μg.The radiochemicalpurity of 125Ⅰ-labeled anti-MIF mAb was still 90.36% after kept at 4℃ for three days.②125I-mMIF mAb could bind to its ligand MIF in vivo with their unaffected immunocompetence and biologic activity.③The lungs,livers,spleens and kidneys of the experimental mice had the relative high radiocounting.CONCLUSION： These results demonstrate that the 125Ⅰ-labeled anti-mMIF mAb made by Iodogen method using the reaction condition introduced in this study had the characteristic including high labeling efficiency,high radiochemicalpurit,good in vitro stability and stable in vivo biologic activity.This study suggests that 125Ⅰ-labeled anti-mMIF mAb had the potential ability to be used in further radioimmunoimaging and targeted therapy research.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2010年第12期1223-1225,共3页
Chinese Journal of Cellular and Molecular Immunology
