頁籤選單縮合
題 名 | 應用聚酯纖維載體培養融合瘤細胞株=Application of Non-Woven Ployester Fabric for Cultivation of Hybridoma Cell Line |
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作 者 | 李文榮; 黃金城; | 書刊名 | 行政院農業委員會家畜衛生試驗所研究報告 |
卷 期 | 41 民95.12 |
頁 次 | 頁153-164 |
分類號 | 437.245 |
關鍵詞 | 不織布聚酯纖維載體; 吊籃式培養瓶; 融合瘤細胞; Non-woven polyester fabric carrier; NWPF carrier; Spinner basket flask; Hybridoma cell; |
語 文 | 中文(Chinese) |
中文摘要 | 現今世界潮流由於動物福祉的意識盛行,以老鼠腹水生產單株抗體的方法並不適合量化生產,對於有大量抗體需求的領域而言,已有許多替代性的方法可供應用。例如以中空纖維(Hollow-fiber)之生物反應器、透析膜(Dialysis membrane)培養瓶製造單株抗體時,分別可以達到0.2~0.3 mg/mL;0.1~1.5 mg/mL抗體濃度,相較於必須不斷繼代培養且分泌抗體濃度僅20 μg/mL的角瓶培養,顯然效能更佳。本試驗以6公克聚酯纖維載體(Polyester fabric carrier)於500 mL吊籃式旋轉瓶進行融合瘤細胞株培養,接種細胞濃度為1x105 cells.mL-1,接種量430 mL。培養期間分別於第5 、6、7、8日以批次式(Batch)各換液約二分之一。自第9日起,採灌注式饋料(Perfusion feed)再繼續培養5日後終止實驗,共收集約4公升培養液。以酵素連結免疫分析法(ELISA)檢測分泌抗體濃度,並與角瓶培養法培養三日以上培養液作對照。各階段收集液抗體反應強度為對照組之2~4倍;總產能大於使用160支T175角瓶每瓶50 mL進行培養後所獲得8公升培養液抗體含量。本試驗細胞株乃分讓而來,無製備所需抗原,因此未定量抗體濃度,惟聚酯纖維載體類似中空纖維,具有高單位體積之表面積比(Surface area/volume ratio)150 cm-1,依結果顯示適合高細胞密度大量培養單融合瘤細胞株。 |
英文摘要 | Monoclonal antibodies(Mabs). have been widely used in biological researches, medical diagnoses and therapies. The production of Mab from ascites by mouse can make a high yield;however, the use of animals for large-scale production of Mab is restricted for the animal welfare concerns. In the current trend, many in vitro alternatives have been developed for Mab production. In this study the hybridoma cell culture was performed in a 500 mL spinner basket containing 6 g non-woven polyester fabric (NWPF) microcarrier. Cells for inoculum were prepared at density 1x105 cell.mL-1 of 430 mL culture medium. Cells were growing in batch culture until day 9 when the perfusion process was initiated. After 5 more days, culture was discontinued. During the 14 days culture period, four liters supernatant was harvested in total. ELISA was applied to evaluate the activity of secreting monoclonal antibody (Mab) from culture . According to the result, the Mab activity from spinner basket culture was 2-4 times higher than T-flask culture in control, and the Mab yield of operation was equivalent to 160 T175 flasks. Application of NWPF material could be a promising alternative to Mab production while optimal culture condition was established. |
本系統中英文摘要資訊取自各篇刊載內容。