Comparison of C-type Nanoantibody Produced in Different Expression Systems Implying Potential Clinical Applications.

IF 4.5 2区 医学 Q2 MEDICINE, RESEARCH & EXPERIMENTAL
Molecular Pharmaceutics Pub Date : 2025-01-06 Epub Date: 2024-12-04 DOI:10.1021/acs.molpharmaceut.4c00980
Moxuan Li, Yancheng Zhan, Bihao Wu, Ye Qin, Jiazhao Gao, Lan Liu, Rui Gong
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Abstract

In the pharmaceutical industry, the Chinese hamster ovary cell, a type of mammalian cell, is extensively employed for the production of conventional full-length monoclonal antibodies. Nanobody is one of the most attractive directions for the development of next-generation antibody drugs. However, a suitable expression system for its manufacture has not yet been comprehensively evaluated. Previously, we proposed that the immunoglobulin constant CH2 domain could be a promising scaffold for developing C-type nanoantibodies (C-Nabs) as candidate therapeutics. Here, we used an antiviral C-Nab, which we identified previously (under review), as a model for investigation. We expressed C-Nabs without a tag in different systems, including a bacterium (C-Nabbac), yeast (C-Nabyeast), and mammalian cell (C-Nabmam). After purification, the binding and neutralizing activities of C-Nabs from different expression systems are similar. Their secondary structures are rich in β-strand. The melting temperatures of C-Nabbac (71.5 °C) and C-Nabmam (70.2 °C) are similar, which are slightly higher than that of C-Nabyeast (65.6 °C), while C-Nabyeast and C-Nabmam are more resistant to urea-induced unfolding than C-Nabbac. C-Nabyeast and C-Nabmam demonstrate higher resistance to aggregation compared to C-Nabbac. C-Nabyeast exhibits greater resistance to enzyme digestion compared to C-Nabbac and C-Nabmam. Notably, when administered via intraperitoneal injection in mice, C-Nabyeast shows superior pharmacokinetics. Overall, after comparing C-Nab proteins from various expression systems, we determined that yeast is the most suitable host for producing C-Nabs. This finding is beneficial for the production of nanobodies as potential drug candidates.

不同表达系统产生的c型纳米抗体的比较暗示潜在的临床应用。
在制药行业,中国仓鼠卵巢细胞是一种哺乳动物细胞,被广泛用于生产传统的全长单克隆抗体。纳米体是新一代抗体药物开发最具吸引力的方向之一。然而,一种适合其制造的表达系统尚未得到全面评价。先前,我们提出免疫球蛋白恒定CH2结构域可能是开发c型纳米抗体(c - nab)的有前途的支架,作为候选治疗药物。在这里,我们使用了一种抗病毒的C-Nab,这是我们之前确定的(正在审查中),作为研究的模型。我们在不同的系统中表达了不带标签的C-Nabs,包括细菌(C-Nabbac)、酵母(C-Nabyeast)和哺乳动物细胞(C-Nabmam)。纯化后,不同表达系统的c - nab的结合和中和活性相似。它们的二级结构中含有丰富的β链。C- nabbac的熔融温度为71.5°C, C- nabmam的熔融温度为70.2°C,略高于C- nabyeast的熔融温度65.6°C, C- nabmam和C- nabmam对尿素诱导展开的抗性强于C- nabbac。与C-Nabbac相比,c - nab酵母和c - nabmann表现出更高的聚集抗性。与C-Nabbac和c - nabmann相比,C-Nabyeast表现出更强的酶消化抗性。值得注意的是,当通过腹腔注射给药小鼠时,C-Nabyeast显示出优越的药代动力学。总的来说,在比较了来自不同表达系统的C-Nab蛋白后,我们确定酵母是最适合产生C-Nab的宿主。这一发现有利于纳米体作为潜在候选药物的生产。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Molecular Pharmaceutics
Molecular Pharmaceutics 医学-药学
CiteScore
8.00
自引率
6.10%
发文量
391
审稿时长
2 months
期刊介绍: Molecular Pharmaceutics publishes the results of original research that contributes significantly to the molecular mechanistic understanding of drug delivery and drug delivery systems. The journal encourages contributions describing research at the interface of drug discovery and drug development. Scientific areas within the scope of the journal include physical and pharmaceutical chemistry, biochemistry and biophysics, molecular and cellular biology, and polymer and materials science as they relate to drug and drug delivery system efficacy. Mechanistic Drug Delivery and Drug Targeting research on modulating activity and efficacy of a drug or drug product is within the scope of Molecular Pharmaceutics. Theoretical and experimental peer-reviewed research articles, communications, reviews, and perspectives are welcomed.
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