Synthesis of pegylated immunonanoparticles
Synthesis of pegylated immunonanoparticles
复制标题
DOI：
10.1023/a:1019842024814
复制
发表时间：
2002-08-01
影响因子：
3.7
通讯作者：
Pardridge, WM
中科院分区：
文献类型：
--
作者：
Olivier, JC;Huertas, R;Pardridge, WM
目的。本文描述了用抗转铁蛋白受体单克隆抗体（MAb）偶联马来酰亚胺接枝聚乙二醇纳米粒子合成聚乙二醇化免疫微粒的方法。聚乳酸（PLA）和双功能聚乙二醇（PEG）制备聚乙二醇化纳米粒子。以八酸锡为催化剂，采用开环聚合法制备了马来酰亚胺- peg (3500)-PLA（40000）和甲氧基peg (2)600-PLA(4)0000共聚物。通过多乳液/溶剂蒸发法制备聚乙二醇化纳米粒子，并通过位于PEG间隔剂末端的马来酰亚胺功能偶联硫代OX26单抗。通过准弹性光散射、凝胶渗透色谱、浊度测定和透射电子显微镜对聚乙二醇化免疫微粒进行了表征。核磁共振波谱证实了这两种共聚物的合成和马来酰亚胺功能的保存。聚乙二醇化免疫微粒的平均直径为121 +/- 5 nm，透射电镜下呈球形。每个聚乙二醇纳米粒子共轭的OX26单抗分子数为67 +/- 4。利用10 nm金与抗小鼠免疫球蛋白二抗的偶联物，在电子显微镜下直接观察到单抗与聚乙二醇化免疫微粒表面的结合。聚乙二醇化免疫粒子可以用双功能聚乙二醇衍生物合成，它可以桥接纳米粒子和靶向单抗。这种新型制剂可以实现小分子、蛋白质药物和基因药物的靶向递送。
Purpose. This work describes the synthesis of pegylated immunonanoparticles by conjugation of an anti-transferrin receptor monoclonal antibody (MAb) to maleimide-grafted pegylated nanoparticles prepared from poly(lactic acid) (PLA) and a bi-functional polyethyleneglycol (PEG).Methods. Maleimide-PEG(3500)-PLA(40000) and methoxyPEG(2)600-PLA(4)0000 copolymers were synthesized by ring opening polymerization of L-lactide using stannous octoate as catalyst. Pegylated nanoparticles were prepared from these copolymers by a multiple emulsion/solvent evaporation method and thiolated OX26 MAb was conjugated through the maleimide function located at the distal end of the PEG spacer. The pegylated immunonanoparticles were characterized by quasi-elastic light scattering, gel permeation chromatography, turbidimetry assays, and transmission electron microscopy.Results. NMR spectroscopy confirmed the synthesis of both copolymers and the preservation of the maleimide function. The pegylated immunonanoparticles had an average diameter of 121 +/- 5 nm and appeared spherical by transmission electron microscopy. The number of OX26 MAb molecules conjugated per individual pegylated nanoparticle was 67 +/- 4. The MAb conjugated to the surface of the pegylated immunonanoparticle was visualized directly by electron microscopy using a conjugate of 10 nm gold and an anti-mouse immunoglobulin secondary antibody.Conclusion. Pegylated immunonanoparticles can be synthesized with bifunctional PEG derivatives that bridge the nanoparticle and the targeting MAb. This novel formulation may enable the targeted delivery of small molecules, protein drugs, and gene medicines.