Department of Chemistry and Center for Cell & Developmental Biology, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, China.
Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Kowloon, Hong Kong SAR, China.
J Control Release. 2024 Jun;370:302-309. doi: 10.1016/j.jconrel.2024.04.038. Epub 2024 May 1.
Displaying antibodies on carrier surfaces facilitates precise targeting and delivery of drugs to diseased cells. Here, we report the synthesis of antibody-lipid conjugates (ALCs) through site-selective acetylation of Lys 248 in human Immunoglobulin G (IgG) and the development of antibody-functionalized red blood cells (immunoRBC) for targeted drug delivery. ImmunoRBC with the HER2-selective antibody trastuzumab displayed on the surface (called Tras-RBC) was constructed following a three-step procedure. First, a peptide-guided, proximity-induced reaction transferred an azidoacetyl group to the ε-amino group of Lys 248 in the Fc domain. Second, the azide-modified IgG was subsequently conjugated with dibenzocyclooctyne (DBCO)-functionalized lipids via strain-promoted azide-alkyne cycloaddition (SPAAC) to result in ALCs. Third, the lipid portion of ALCs was then inserted into the cell membranes, and IgGs were displayed on red blood cells (RBCs) to construct immunoRBCs. We then loaded Tras-RBC with a photosensitizer (PS), Zinc phthalocyanine (ZnPc), to selectively target HER2-overexpressing cells, release ZnPc into cancer cells following photolysis, and induce photodynamic cytotoxicity in the cancer cells. This work showcases assembling immunoRBCs following site-selective lipid conjugation on therapeutic antibodies and the targeted introduction of PS into cancer cells. This method could apply to the surface functionalization of other membrane-bound vesicles or lipid nanoparticles for antibody-directed drug delivery.
将抗体展示在载体表面上有助于将药物精确靶向并递送到病变细胞。在这里，我们报告了通过赖氨酸 248 在人免疫球蛋白 G（IgG）中的位点选择性乙酰化合成抗体-脂质缀合物（ALC），并开发了用于靶向药物递送的抗体功能化红细胞（immunoRBC）。表面展示 HER2 选择性抗体曲妥珠单抗的 immunoRBC（称为 Tras-RBC）是通过三步程序构建的。首先，肽引导的近程诱导反应将叠氮乙酰基转移到 Fc 结构域中赖氨酸 248 的 ε-氨基上。其次，通过应变促进的叠氮化物-炔烃环加成（SPAAC）将叠氮化物修饰的 IgG 与二苯并环辛炔（DBCO）功能化脂质缀合，得到 ALC。第三，ALC 的脂质部分然后插入细胞膜中，IgG 展示在红细胞（RBC）上构建 immunoRBC。然后，我们将光敏剂（PS）锌酞菁（ZnPc）加载到 Tras-RBC 中，以选择性靶向过表达 HER2 的细胞，在光解后将 ZnPc 释放到癌细胞中，并在癌细胞中诱导光动力细胞毒性。这项工作展示了在治疗性抗体上进行位点选择性脂质缀合后组装 immunoRBC，并将 PS 靶向引入癌细胞。这种方法可应用于其他膜结合囊泡或脂质纳米颗粒的表面功能化，以进行抗体导向的药物递送。
