抗体可以作为半抗原二硝基氟苯免疫靶向治疗的载体  

A potential carrier for targeted therapy of DNFB hapten-induced immune

作　　者：何晓俊[1] 储艳秋[2] 蒋公羽[2] 戴兆云[3] 

机构地区：[1]金坛市人民医院内科,江苏省金坛市214300 [2]复旦大学化学系,上海市200433 [3]复旦大学附属华东医院消化科,上海市200040

出　　处：《世界华人消化杂志》2009年第13期1350-1354,共5页World Chinese Journal of Digestology

基　　金：上海市中医肝病基金资助项目;No.SH-ZXYJH2005-004~~

摘　　要：目的:了解半抗原二硝基氟苯(DNFB)结合乙型肝炎免疫球蛋白(HBIG)后对其抗体功能的影响.方法:常温下把HBIG(13mg/500μLPBS)水溶液分别与不同剂量(2、4、8mg)的DNFB混合,组成2、4、8mg管,混匀过夜后离心.取2、4mg管及未加DNFB的HBIG水溶液对照管,各取半量对半稀释,以HiTrap分子筛柱脱盐,3个洗脱液再分别作对半系列稀释11次,3组共组成36个标本.ELISA法检测乙肝表面抗体的效价.取2、4mg管的另一半分别予饱和硫酸铵沉淀后取上清,以紫外光光度分析仪测定A340,与加入不同剂量DNFB的5个系列浓度作内标的对照管上清相比较,计算相应管中HBIG结合掉的DNFB量.结果:2、4mg管及对照组,经双因素方差分析及q检验,三组间同稀释度间吸光差值(A450-A630)无差异,而各稀释度间则有非常显著差异(P<0.01).饱和硫酸铵沉淀结合分光光度法测定证实2、4mg管中6.5mgHBIG分别结合DNFB0.67mg和0.86mg,即每一个HBIG分子分别结合88.72及113.48个DNFB半抗原分子,约占HBIG氨基酸残基的6.12%和7.86%.结论:免疫球蛋白(抗体)可作为半抗原DNFB免疫靶向治疗慢性病毒感染或肿瘤的一种有效载体.AIM： To investigate the influence of dinitroflourobenzene （DNFB） on the antibody titer of hepatitis B immunoglobulin （HBIG）. METHODS： Three different doses （2, 4, 8 mg） of DNFB were mixed with phosphate buffer solution （PBS） of HBIG respectively, 13 mg in a volume of 500 μL PBS, while same solutions of HBIG were used as controls. The 2, 4 mg tubes and the control tubes were not precipitated except for the 8 mg tube with 10 000 r/min config- uration. The 3 transparent tubes were desalted with column of Sephadex G-25 Superfine HiTrap respectively. All of the eluents of the 3 tubes were half-diluted consecutively with phosphate buffer saline （PBS） respectively to change totally into 36 samples of 2 mg group, 4 mg group and control group, with 12 in each group. The above samples were all tested for the antibody fiter of HBIG using ELISA. The 2 mg tube, 4 mg tube and the control tube were all precipitated in solution of saturated （NH4）2SO4. Compared with standard curves of both quantity of DNFB and gauges of A340 in supernants made of same controlled tubes, the remaining DNFB in supernants in different tubes were all obtained respectively. RESULTS： Except for the 8 mg group, there were no difference in the 3 groups in terms of the absorbance values （A450-A630） of HBIG in same dilution grade of any group with two-way anova （analysis of variance）（P 〉 0.05）. However there was great difference between different dilution grades in any group with two-way anova （P 〈 0.01）. The 6.5 mg of HBIG in the 2 mg tube and 4 mg tube were respectively combined with 0.67 mg and 0.86 mg of DNFB, meaning that each molecule of HBIG with 88.72 and 103.48 molecules of DNFB, namely 6.12% and 7.86% in residual amino acid of each molecule of HBIG combined with DNFB. CONCLUSION： The immune globulin （antibody） might be used as a carrier for DNFB target therapy.

关 键 词：乙型肝炎免疫球蛋白 二硝基氟苯 靶向治疗 抗HBS 半抗原 

分 类 号：R392[医药卫生—免疫学]