> High capacity magnetic protein A agarose beads, LOABeads PrtA, were used in the development of a new process for affinity purification of monoc'/> Pilot‐scale process for magnetic bead purification of antibodies directly from non‐clarified CHO cell culture
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Pilot‐scale process for magnetic bead purification of antibodies directly from non‐clarified CHO cell culture

机译：直接来自非澄清Cho细胞培养的磁珠净化抗体的试验规模过程

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> High capacity magnetic protein A agarose beads, LOABeads PrtA, were used in the development of a new process for affinity purification of monoclonal antibodies (mAbs) from non‐clarified CHO cell broth using a pilot‐scale magnetic separator. The LOABeads had a maximum binding capacity of 65?mg/mL and an adsorption capacity of 25–42?mg IgG/mL bead in suspension for an IgG concentration of 1 to 8?g/L. Pilot‐scale separation was initially tested in a mAb capture step from 26?L clarified harvest. Small‐scale experiments showed that similar mAb adsorptions were obtained in cell broth containing 40?×?10 6 cells/mL as in clarified supernatant. Two pilot‐scale purification runs were then performed on non‐clarified cell broth from fed‐batch runs of 16?L, where a rapid mAb adsorption ≥96.6% was observed after 1?h. This process using 1 L of magnetic beads had an overall mAb yield of 86% and 16 times concentration factor. After this single protein A capture step, the mAb purity was similar to the one obtained by column chromatography, while the host cell protein content was very low, 10 ppm. Our results showed that this magnetic bead mAb purification process, using a dedicated pilot‐scale separation device, was a highly efficient single step, which directly connected the culture to the downstream process without cell clarification. Purification of mAb directly from non‐clarified cell broth without cell separation can provide significant savings in terms of resources, operation time, and equipment, compared to legacy procedure of cell separation followed by column chromatography step. ? 2019 American Institute of Chemical Engineers Biotechnol. Prog ., 35: e2775, 2019.
机译： > 高容量磁性蛋白A琼脂糖珠子，LOABEADS PRTA，用于使用先导级磁分离器从非澄清CHO细胞肉汤的亲和纯化单克隆抗体（mAb）的新方法。 LOABEADS的最大粘合能力为65Ωmg/ ml，吸附容量为25-42×mg IgG / ml胎珠，其IgG浓度为1至8Ω·克/升。最初在MAB捕获步骤中最初在26μl澄清的收获中进行试验级分离。小规模实验表明，在含有40Ω·汤的细胞肉汤中获得了类似的MAB吸附 6 细胞/ mL如澄清的上清液中。然后在来自16·L的FED-批量运行的非澄清细胞肉汤上进行两种先导级净化运行，其中在1℃后观察到快速mAb吸附≥96.6％。使用1L磁珠的该方法总体mAb产率为86％和16次浓度因子。在该单蛋白捕获步骤之后，MAB纯度类似于通过柱色谱所获得的纯度，而宿主细胞蛋白质含量非常低，10 ppm。我们的研究结果表明，该磁珠MAB净化工艺使用专用的先导级分离装置是一种高效的单步，其直接将培养物连接到下游工艺而无需细胞澄清。与无细胞分离的非澄清细胞肉汤直接从未澄清的细胞肉汤纯化可以在资源，操作时间和设备方面提供显着的节省，与细胞分离的传统程序相比，柱色谱步骤。还2019年美国化学工程研究所 生物技术。 PROG 。，35：E2775，2019。

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