製備磁化竹炭發展高效率的細胞篩選器應用於單株抗體生產之研究

單株抗體的製備在1975年由George Kohler和Cesar Milstein成功地利用細胞融合的技術製造出來後就開始被廣泛的運用，然而其在操作流程上較為繁瑣，尤其在選殖融合瘤細胞(hybridoma cell)單株化的過程時，不僅需要花費大量的時間、步驟繁瑣、同時也必須消耗很多的人力，且操作者的熟練度往往對結果影響很大，由於以上諸多缺點，使單株抗體的製備一直無法有效地大量生產。先前，在本實驗室中，我們開發出一組高產量的細胞篩選器，成功的縮短單株化選殖的時間，此種細胞篩選器是利用本實驗室開發出來的虹吸式生物反應器(cytoflow-bioreactor)配合竹炭作為載體(carrier)。但是目前此細胞篩選器還是有一些問題，如載體(carrier)的顆粒大小受到限制、非專一性吸附的問題嚴重、專一性鍵結(specific binding)上carrier的細胞數量太少、專一性融合瘤細胞(specific hybridoma cell)的生產效率差以及無法有效地分離carrier和細胞…等等問題。因此，本研究主要目的在於希望能改善此細胞篩選器(Cytoflow-Bioreactor base Cell Sorter, CBCS)的種種問題，提高細胞篩選器的分選效率與專一性融合瘤細胞(specific hybridoma cell)的生產效率，同時避免非專一性鍵結(non-specific binding)的可能性和降低操作上的困難，發展出高效率的虹吸式細胞篩選器(High-efficiency Cytoflow-Bioreactor base Cell Sorter, HCBCS)，最終達到單株抗體大量生產的目的。

關鍵字

生物反應器；融合瘤；虹吸式反應器；單株抗體；磁化竹炭；表面改質

並列摘要

George Kohler and Cesar Milstein employed fusion theory to produce monoclonal antibody in 1975. Monoclonal antibodies (mAbs) have been proven useful in research and clinical applications. However, the generation of mAbs by conventional hybridoma technology is time-, cost- and labor-consuming. Before, a siphon bioreactor and the carrier(bamboo charcoal) combined as a cell sorter called : Cytoflow-bioreactor base cell sorter(CBCS). Our design allows the production of mAbs while avoiding time-consuming steps, such as large numbers of limiting dilutions and screening assays, and demonstrates that the CBCS could be a powerful tool for monoclonal antibody production. But the CBCS still had some problems : the particle size of carrier is limited、non-specific binding、low-efficiency cell sorting and operational difficulties. The purpose of study is improving cytoflow-bioreactor base cell sorter, promote cell sorting efficiency, avoid the possibility of non-specific binding and operational difficulties, create a high-efficiency cytoflow-bioreactor base cell sorter(HCBCS) could be employed to harvest antibody-secreting B cells from the peripheral blood without sacrificing the animal. In the future, we hope this method not only produce animals monoclonal antibody but also produce humanized monoclonal antibody, to achieve the purpose of monoclonal antibody mass production.

並列關鍵字

Bioreactor ； Hybridomas ； Cytoflow reactor ； Monoclonal antibody ； Magnetic Bamboo charcoal ； Surface modification

參考文獻

1. Kern, D. and E.R.P. Zuiderweg, The role of dynamics in allosteric regulation. Current Opinion in Structural Biology, 2003. 13(6): p. 748-757.

2. Cantrell, D., T cell antigen receptor signal transduction pathways. Annual Review of Immunology, 1996. 14: p. 259-274.

5. Zhang, H.T., et al., ErbB receptors: from oncogenes to targeted cancer therapies. Journal of Clinical Investigation, 2007. 117(8): p. 2051-2058.

6. Goodey, N.M. and S.J. Benkovic, Allosteric regulation and catalysis emerge via a common route. Nature Chemical Biology, 2008. 4(8): p. 474-482.

7. Kohler, G. and C. Milstein, Continuous Cultures of Fused Cells Secreting Antibody of Predefined Specificity. Nature, 1975. 256(5517): p. 495-497.

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製備磁化竹炭發展高效率的細胞篩選器應用於單株抗體生產之研究

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