Centre for Bioseparation Technology, Vellore Institute of Technology (VIT), 632014 Vellore, Tamilnadu, India.
Front Biosci (Elite Ed). 2024 Aug 7;16(3):26. doi: 10.31083/j.fbe1603026.
Monoclonal antibodies (mAbs) are pioneers in the diagnosis and treatment of many diseases, such as cancer, asthma, poisoning, viral infections, etc. As the market value of mAbs increases in the biopharma industry, the demand for high quantities is met by upscaled production using bioreactor systems. Thus, disposable, porous matrices called cryogels have gained the primary focus for adherent support in the proliferation of hybridoma cells.
In this study, a gelatin-immobilized polyhydroxyethylmethacrylate-based cryogel material (disc-shaped, 9 mL bed volume) was synthesized, and a mini-bioreactor set up developed for culturing hybridoma cells to produce mAbs continuously. The hybridoma clone, 1B4A2D5, secreting anti-human serum albumin monoclonal antibodies, was immobilized in the cryogel matrix (2 discs, 18 mL bed volume).
The hybridoma cells were attached to the matrix within 12 h after inoculation, and the cells were in the lag phase for seven days, where they were secreted mAb into the circulation medium. During the initial exponential phase, the glucose consumption, lactic acid production, and mAb production were 3.36 mM/day, 3.67 mM/day, and 55.61 µg/mL/day, respectively. The medium was refreshed whenever the glucose in the media went below 50% of the initial glucose concentration. The cryogenic reactor was run continuously for 25 days, and the mAb concentration reached a maximum on the 17th day at 310.59 µg/mL.
The cumulative amount of mAbs produced in 25 days of running was 246 µg/mL, 7.7 times higher than the mAbs produced from T-flask batch cultivation. These results demonstrate that the developed polyhydroxyethylmethacrylate-based cryogel reactor can be used efficiently for continuous mAb production.
单克隆抗体（mAbs）是许多疾病诊断和治疗的先驱，如癌症、哮喘、中毒、病毒感染等。随着生物制药行业中 mAbs 的市场价值不断增加，对大量 mAbs 的需求通过使用生物反应器系统进行规模化生产来满足。因此，用于支持杂交瘤细胞增殖的贴壁支撑的一次性、多孔基质称为冷冻凝胶得到了主要关注。
在这项研究中，合成了一种明胶固定的聚羟乙基甲基丙烯酸酯基冷冻凝胶材料（圆盘状，9 毫升床体积），并开发了一个小型生物反应器系统，用于连续培养杂交瘤细胞以生产 mAbs。分泌抗人血清白蛋白单克隆抗体的杂交瘤克隆 1B4A2D5 被固定在冷冻凝胶基质中（2 个圆盘，18 毫升床体积）。
杂交瘤细胞在接种后 12 小时内附着在基质上，细胞在 7 天的迟滞期内，将 mAb 分泌到循环培养基中。在初始指数生长期，葡萄糖消耗、乳酸生成和 mAb 生产分别为 3.36 mM/天、3.67 mM/天和 55.61 µg/mL/天。当培养基中的葡萄糖下降到初始葡萄糖浓度的 50%以下时，就会更换新鲜培养基。冷冻反应器连续运行 25 天，mAb 浓度在第 17 天达到 310.59 µg/mL 的最大值。
在 25 天的运行中，共生产了 246 µg/mL 的 mAb，是 T 瓶分批培养生产 mAb 的 7.7 倍。这些结果表明，开发的聚羟乙基甲基丙烯酸酯基冷冻凝胶反应器可有效地用于连续 mAb 生产。
