Sinegubova Maria, Vorobiev Ivan, Klishin Anatoly, Eremin Dmitry, Orlova Nadezhda, Orlova Natalya, Polzikov Mikhail
Laboratory of Mammalian Cell Bioengineering, Institute of Bioengineering, Federal State Institution «Federal Research Centre «Fundamentals of Biotechnology» of the Russian Academy of Sciences», Leninsky Prospect, 33, Build. 2, 119071 Moscow, Russia.
IVFarma LLC, Nauchnyi Proezd, 20, Build. 2, 117246 Moscow, Russia.
Pharmaceutics. 2022 Jan 1;14(1):96. doi: 10.3390/pharmaceutics14010096.
Recombinant human follicle stimulating hormone (r-hFSH) is widely used for infertility treatment and is subject to the development of biosimilars. There are different purification strategies that can yield r-hFSH of pharmaceutical quality from Chinese hamster ovary cell culture broth. We developed a purification process for r-hFSH centered on immunoaffinity chromatography with single-domain recombinant camelid antibodies. The resulting downstream process is simple and devoid of ultrafiltration operations. Studies on chromatography resin resource and ligand leakage showed that the immunoaffinity matrix employed was suitable for industrial use and stable for at least 40 full chromatography cycles, and the leaked single-domain antibody ligand was completely removed by subsequent purification steps. All chromatography resins employed withstood the same 40 cycles of use without significant changes in separation efficiency and product binding capacity. The resulting industrial purification process yielded batches of r-hFSH with consistent levels of purity and bioactivity.
重组人促卵泡激素（r-hFSH）广泛用于不孕症治疗，并且面临生物类似药的开发。有不同的纯化策略可从中国仓鼠卵巢细胞培养液中获得具有药物质量的r-hFSH。我们开发了一种以单域重组骆驼科抗体免疫亲和色谱为核心的r-hFSH纯化工艺。由此产生的下游工艺简单，且无需超滤操作。对色谱树脂资源和配体泄漏的研究表明，所使用的免疫亲和基质适用于工业用途，并且至少在40个完整的色谱循环中稳定，后续纯化步骤可完全去除泄漏的单域抗体配体。所有使用的色谱树脂都经受了相同的40个循环的使用，分离效率和产品结合能力没有显著变化。由此产生的工业纯化工艺生产出了纯度和生物活性水平一致的r-hFSH批次。
