Beyond affinity: selection of antibody variants with optimal biophysical properties and reduced immunogenicity from mammalian display libraries.
Beyond affinity: selection of antibody variants with optimal biophysical properties and reduced immunogenicity from mammalian display libraries.
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DOI：
10.1080/19420862.2020.1829335
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发表时间：
2020-01
期刊：
影响因子：
5.3
通讯作者：
McCafferty J
中科院分区：
文献类型：
--
作者：
Dyson MR;Masters E;Pazeraitis D;Perera RL;Syrjanen JL;Surade S;Thorsteinson N;Parthiban K;Jones PC;Sattar M;Wozniak-Knopp G;Rueker F;Leah R;McCafferty J
蛋白质药物开发的早期阶段传统上集中于导致期望的治疗作用模式的靶标结合特性。随着越来越多的蛋白质治疗剂通过开发管道;然而，很明显，可能会出现非最佳的生物物理特性，特别是当蛋白质以高浓度配制时，导致聚集或多反应性。这种后期的“可开发性”问题可能导致开发过程的延迟或失败。聚集倾向也与增加的免疫原性相关，导致昂贵的晚期临床失败。使用核酸酶定向整合，我们构建了大型哺乳动物展示文库，其中每个细胞含有插入在单个位点的单个抗体基因/细胞，从而实现转录标准化。我们显示了在哺乳动物细胞中实现的差的生物物理特性和展示水平之间的强相关性，这不是通过酵母展示复制的。使用两个充分记录的具有差的生物物理特征的抗体的实例（MEDI-1912和bococizumab），基于表面疏水性和正电荷补丁创建变体文库。使用哺乳动物展示来选择保留靶结合并允许增加的展示水平的抗体。所得变体表现出降低的多反应性和降低的聚集倾向。此外，我们在bococizumab的情况下表明，生物免疫学改进的变体比亲本分子的免疫原性更低。因此，哺乳动物展示有助于解决铅发现的最早阶段的多个可开发性问题，从而显著降低蛋白质药物未来开发的风险。
The early phase of protein drug development has traditionally focused on target binding properties leading to a desired mode of therapeutic action. As more protein therapeutics pass through the development pipeline; however, it is clear that non-optimal biophysical properties can emerge, particularly as proteins are formulated at high concentrations, causing aggregation or polyreactivity. Such late-stage “developability” problems can lead to delay or failure in traversing the development process. Aggregation propensity is also correlated with increased immunogenicity, resulting in expensive, late-stage clinical failures. Using nucleases-directed integration, we have constructed large mammalian display libraries where each cell contains a single antibody gene/cell inserted at a single locus, thereby achieving transcriptional normalization. We show a strong correlation between poor biophysical properties and display level achieved in mammalian cells, which is not replicated by yeast display. Using two well-documented examples of antibodies with poor biophysical characteristics (MEDI-1912 and bococizumab), a library of variants was created based on surface hydrophobic and positive charge patches. Mammalian display was used to select for antibodies that retained target binding and permitted increased display level. The resultant variants exhibited reduced polyreactivity and reduced aggregation propensity. Furthermore, we show in the case of bococizumab that biophysically improved variants are less immunogenic than the parental molecule. Thus, mammalian display helps to address multiple developability issues during the earliest stages of lead discovery, thereby significantly de-risking the future development of protein drugs.
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10.4
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Sette A