用于免疫脂质体和双特异性抗体复合物的人免疫球蛋白 G 的位点选择性赖氨酸乙酰化,Journal of the American Chemical Society

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用于免疫脂质体和双特异性抗体复合物的人免疫球蛋白 G 的位点选择性赖氨酸乙酰化
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2022-09-27 , DOI: 10.1021/jacs.2c07594
Dingdong Yuan ₁ , Yu Zhang ₁ , King Hoo Lim ₂ , Stephen King Pong Leung ₃ , Xizi Yang ₃ , Yujie Liang ₁ , Wilson Chun Yu Lau ₃ , Kwan T Chow ₂ , Jiang Xia ₁

Affiliation

蛋白质中单个赖氨酸残基的位点选择性乙酰化达到赖氨酸乙酰转移酶的准确性、精密度和可靠性具有挑战性。在这里，我们报告了一种肽引导、邻近驱动的基团转移反应，该反应使人免疫球蛋白 G (IgG) 重链中片段可结晶区 (Fc 区) 的单个赖氨酸残基 Lys 248 乙酰化。与 Fc 结构域结合的 Fc 相互作用肽将酚酯定位在靠近 Lys 248 的位置，这会诱导亲核反应并导致乙酰基转移到 Lys 248。乙酰化反应在生理条件下进行到可观的产率无需去糖基化、非天然氨基酸或催化剂。除了乙酰化，功能部分如叠氮化物、炔烃、荧光分子、或生物素也可以选择性地安装在 Lys 248 上，允许 IgG 进一步衍生化。然后，我们合成了一种抗体-脂质偶联物并构建了抗体偶联脂质体（免疫脂质体），靶向 HER2 阳性 (HER2+) 癌细胞。我们还构建了一个双特异性抗体复合物 (bsAbC)，共价连接抗 HER2 抗体和抗 CD3 抗体。bsAbC 显示纳摩尔浓度的体外效应细胞介导的细胞毒性。与双特异性抗体 (bsAbs) 相比，bsAbCs 是基于天然 IgG 构建的，每个抗原包含两个抗原结合位点，是 bsAbs 的两倍。总之，这项工作报告了一种对天然抗体进行位点选择性乙酰化的方法，强调了一种简便的位点选择性 IgG 功能化方法，并强调了 bsAbC 在癌症免疫治疗中的潜力。

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Site-Selective Lysine Acetylation of Human Immunoglobulin G for Immunoliposomes and Bispecific Antibody Complexes

Site-selective acetylation of a single lysine residue in a protein that reaches a lysine acetyltransferase’s accuracy, precision, and reliability is challenging. Here, we report a peptide-guided, proximity-driven group transfer reaction that acetylates a single lysine residue, Lys 248, of the fragment crystallizable region (Fc region) in the heavy chain of the human Immunoglobulin G (IgG). An Fc-interacting peptide bound with the Fc domain and positioned a phenolic ester close to Lys 248, which induced a nucleophilic reaction and resulted in the transfer of an acetyl group to Lys 248. The acetylation reaction proceeded to a decent yield under the physiological condition without the need for deglycosylation, unnatural amino acids, or catalysts. Along with acetylation, functional moieties such as azide, alkyne, fluorescent molecules, or biotin could also be site-selectively installed on Lys 248, allowing IgG’s further derivatization. We then synthesized an antibody–lipid conjugate and constructed antibody-conjugated liposomes (immunoliposomes), targeting HER2-positive (HER2+) cancer cells. We also built a bispecific antibody complex (bsAbC) covalently linking an anti-HER2 antibody and an anti-CD3 antibody. The bsAbC showed in vitro effector-cell-mediated cytotoxicity at nanomolar concentrations. Compared with bispecific antibodies (bsAbs), bsAbCs are constructed based on native IgGs and contain two antigen-binding sites to each antigen, twice that of bsAbs. Altogether, this work reports a method of site-selective acetylation of native antibodies, highlights a facile way of site-selective IgG functionalization, and underscores the potential of bsAbCs in cancer immunotherapy.

更新日期：2022-09-27

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