MOE Key Laboratory of Smart Drug Delivery, School of Pharmacy & State Key Laboratory of Molecular Engineering of Polymers, Fudan University, Shanghai, 201203 P.R. China.
Department of Pharmacology, School of Basic Medical Sciences & Center of Medical Research and Innovation, Shanghai Pudong Hospital, Fudan University, Shanghai, 200032 P.R. China.
Nano Lett. 2021 Dec 8;21(23):10107-10113. doi: 10.1021/acs.nanolett.1c03946. Epub 2021 Nov 23.
PEGylated nanocarriers have gained increasing attention due to reduced toxicity and enhanced circulation compared with free drugs. According to guidances of drug regulatory departments worldwide, it is crucial to determine free and liposomal drug concentrations; however, the conventional used separation methods including dialysis, ultrafiltration, and solid-phase extraction (SPE) have drawbacks of time-consuming, drug leakage, environmental pollution or error bias of trace level drug. Here we developed a facile PEG-scFv-based separation method combined with HPLC to quantify free doxorubicin (DOX) and liposomal DOX in plasma. Anti-PEG single chain variable fragment antibody (PEG-scFv) was adopted to sediment PEGylated liposomes by simple incubation and low speed centrifugation. Compared to SPE, it demonstrated sufficient accuracy and sensitivity to evaluate free and liposomal DOX with intact liposomes. Therefore, it can serve as an alternative approach of SPE, which is suitable for quality assessment and pharmacokinetics evaluation of PEGylated liposomal drugs and possible other PEGylated nanocarriers.
聚乙二醇化纳米载体由于毒性降低和循环时间延长而受到越来越多的关注。根据全球药物监管部门的指导，确定游离药物和脂质体药物的浓度至关重要；然而，传统的分离方法包括透析、超滤和固相萃取（SPE）都存在耗时、药物泄漏、环境污染或痕量药物的误差偏差等缺点。在这里，我们开发了一种简单的基于 PEG-scFv 的分离方法，结合 HPLC 定量检测血浆中的游离阿霉素（DOX）和脂质体 DOX。采用抗聚乙二醇单链可变片段抗体（PEG-scFv）通过简单孵育和低速离心沉淀聚乙二醇化脂质体。与 SPE 相比，该方法具有足够的准确性和灵敏度，可用于评估完整脂质体的游离和脂质体 DOX。因此，它可以作为 SPE 的替代方法，适用于聚乙二醇化脂质体药物和可能的其他聚乙二醇化纳米载体的质量评估和药代动力学评价。
