Production of antibodies and antibody fragments containing non-natural amino acids in Escherichia coli.

IF 5.6 2区 医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL
mAbs Pub Date : 2024-01-01 Epub Date: 2024-02-21 DOI:10.1080/19420862.2024.2316872
Jacquelyn Blake-Hedges, Dan Groff, Wilson Foo, Jeffrey Hanson, Elenor Castillo, Miao Wen, Diana Cheung, Mary Rose Masikat, Jian Lu, Young Park, Nina Abi Carlos, Hans Usman, Kevin Fong, Abigail Yu, Sihong Zhou, Joyce Kwong, Cuong Tran, Xiaofan Li, Dawei Yuan, Trevor Hallam, Gang Yin
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引用次数: 0

Abstract

Therapeutic bioconjugates are emerging as an essential tool to combat human disease. Site-specific conjugation technologies are widely recognized as the optimal approach for producing homogeneous drug products. Non-natural amino acid (nnAA) incorporation allows the introduction of bioconjugation handles at genetically defined locations. Escherichia coli (E. coli) is a facile host for therapeutic nnAA protein synthesis because it can stably replicate plasmids encoding genes for product and nnAA incorporation. Here, we demonstrate that by engineering E. coli to incorporate high levels of nnAAs, it is feasible to produce nnAA-containing antibody fragments and full-length immunoglobulin Gs (IgGs) in the cytoplasm of E. coli. Using high-density fermentation, it was possible to produce both of these types of molecules with site-specifically incorporated nnAAs at titers > 1 g/L. We anticipate this strategy will help simplify the production and manufacture of promising antibody therapeutics.

在大肠杆菌中生产含有非天然氨基酸的抗体和抗体片段。
治疗性生物共轭物正在成为对抗人类疾病的重要工具。位点特异性共轭技术被广泛认为是生产均一药物产品的最佳方法。非天然氨基酸(nnAA)掺入技术可在基因定义的位置引入生物共轭手柄。大肠杆菌(E. coli)是治疗性 nnAA 蛋白合成的理想宿主,因为它能稳定复制编码产品和 nnAA 结合基因的质粒。在这里,我们证明,通过对大肠杆菌进行工程改造,使其能够结合高水平的 nnAA,就可以在大肠杆菌的细胞质中生产含有 nnAA 的抗体片段和全长免疫球蛋白 G(IgG)。利用高密度发酵法,我们可以生产出这两类分子,并在滴度大于 1 克/升时特异性地结合了 nnAAs。我们预计这种策略将有助于简化有前景的抗体疗法的生产和制造。
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
mAbs
mAbs 工程技术-仪器仪表
CiteScore
10.70
自引率
11.30%
发文量
77
审稿时长
6-12 weeks
期刊介绍: mAbs is a multi-disciplinary journal dedicated to the art and science of antibody research and development. The journal has a strong scientific and medical focus, but also strives to serve a broader readership. The articles are thus of interest to scientists, clinical researchers, and physicians, as well as the wider mAb community, including our readers involved in technology transfer, legal issues, investment, strategic planning and the regulation of therapeutics.
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