基于人类单克隆抗体异体混合物的实验性蛇毒抗血清的单批次表达

Single-Batch Expression of an Experimental Recombinant Snakebite Antivenom Based on an Oligoclonal Mixture of Human Monoclonal Antibodies

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Biotechnology Journal

影响因子:3.1

分区:生物学3区 / 生化研究方法3区生物工程与应用微生物3区

发表日期:2024 Oct

作者: Anna C Adams, Lise M Grav, Shirin Ahmadi, Camilla Holst Dahl, Anne Ljungars, Andreas H Laustsen, Lars K Nielsen

DOI: 10.1002/biot.202400348

摘要

异体单克隆抗体（oligoclonal antibodies）是由精心定义的多克隆抗体混合物，广泛用于治疗复杂疾病，如感染和癌症。除了这些医学领域外，它们还可以用于蛇毒毒血清的治疗，重组生产的人源单克隆抗体能够克服传统抗血清的许多缺陷。然而，在工业生产中制造多个单克隆抗体批次的成本很高。因此，将多个抗体生产的细胞系在单一批次中混合，形成异体抗体，成为一种具有吸引力的策略，以实现仅有一个上游和下游工艺。本研究选择了四种针对不同毒素的抗体，这些毒素存在于多种眼镜蛇属和眼镜蛇科蛇类毒液中，构建了稳定的抗体生产细胞系。共培养后，发现细胞系比例在7天内保持稳定。纯化的异体抗体鸡尾酒中含有预期的抗体浓度，并能按预期结合目标毒素。这些结果为在单一批次中混合多细胞系以重组制造定制抗蛇毒血清的策略提供了可行性验证，未来可应用于蛇咬毒的治疗及其他可能需要异体抗体混合物的领域。

Abstract

Oligoclonal antibodies, which are carefully defined mixtures of monoclonal antibodies, are valuable for the treatment of complex diseases, such as infectionss and cancer. In addition to these areas of medicine, they could be utilized for the treatment of snakebite envenoming, where recombinantly produced monoclonal human antibodies could overcome many of the drawbacks accompanying traditional antivenoms. However, producing multiple individual batches of monoclonal antibodies in an industrial setting is associated with significant costs. Therefore, it is attractive to produce oligoclonal antibodies by mixing multiple antibody-producing cell lines in a single batch to have only one upstream and downstream process. In this study, we selected four antibodies that target different toxins found in the venoms of various elapid snake species, such as mambas and cobras, and generated stable antibody-producing cell lines. Upon co-cultivation, we found the cell line ratios to be stable over 7 days. The purified oligoclonal antibody cocktail contained the anticipated antibody concentrations and bound to the target toxins as expected. These results thus provide a proof of concept for the strategy of mixing multiple cell lines in a single batch to manufacture tailored antivenoms recombinantly, which could be utilized for the treatment of snakebite envenoming and in other fields where oligoclonal antibody mixtures could find utility.