頂尖細胞療法公司都點頭的技術規格

諾華(Novartis)、凱德(Kite Pharma)、朱諾(Juno)是免疫細胞療法的代表性公司。其中,諾華及凱德各自研發的Kymriah、Yescarta是目前全球唯二獲得FDA批准上市的CAR-T治療產品,朱諾則正積極爭取成為第三家。世界上研究CAR-T的生技公司不在少數,何以它們能率先進入市場,三者間是否有共通點可供大家借鑒?

從研究走向市場
當細胞治療從臨床走向商品化,除了考量法規問題外,如何優化製程得以量產並輸出穩定品質是產品能否成功的重要因素。

檢視三家公司的專利文件、文獻資料可以發現製程優化的方向一致在於簡化流程、導入自動化、縮短生產時間。值得注意的是諾華、凱德及朱諾無獨有偶都在製程裡導入了一項設備 – Sepax細胞血球分離機作為優化的一環顯示其重要性。以下說明諾華、凱德及朱諾在各自製程中是如何使用Sepax細胞血球分離機

諾華、凱德及朱諾細胞治療製程都涉及使用相同的設備-Sepax細胞血球分離機

諾華製程
諾華導入Sepax後主要運用在細胞洗滌以及密度梯度離心分離取得MNC。下面擷取自諾華的CAR-T製程專利文件中的一段,其描述Sepax用於洗滌回溫後的白血球濃厚液移除細胞碎片、游離血紅素與抗凝劑以及搭配介質OptiPrep進行密度梯度分離取得MNC:

The Sepax 2 RM device (Biosafe) was used to wash thawed leukopak cells to remove subcellular debris, free hemoglobin and cryoprotectants, to achieve volume reduction, and to enable subsequent Ficoll density gradient separation.

……….略

Operators performed the Sepax 2 NeatCell protocol and where the protocol called for Ficoll, it was replaced with OptiPrep. FIG. 18 shows a schematic on how the Sepax 2 NeatCell single use disposable tubing kit (Biosafe CS900.2) is configured with the cell and medium bags.

Novartis Ag, Methods of making chimeric antigen receptor -expressing cells, WO/2017/117112, 2017-07-06

凱德製程
凱德導入Sepax後主要運用在細胞減量密度梯度離心分離以及細胞洗滌。下圖左側為美國癌症研究所(National Cancer Institute, NCI)開發的CAR-T流程,右側為經凱德優化後的流程,優化後的流程以Sepax取代手動Ficol離心、細胞洗滌(活化後、培養後),比起原先製程更具封閉性縮短生產期

Lu,TangyingLilyet al. “A Rapid Cell Expansion Process for Production of Engineered Autologous CAR-T Cell Therapies.” Human gene therapy methods vol. 27,6 (2016): 209-218. doi:10.1089/hgtb.2016.120

朱諾製程
朱諾導入Sepax後主要用在細胞洗滌磁珠結合。下面擷取自朱諾的CAR-T製程專利文件中的一段,其描述使用Sepax洗滌白血球濃厚液(去血小板、培養液)再藉由Sepax進行細胞、磁珠結合的過程:

Cells of the leukapheresis sample were washed and resuspended in a selection buffer for affinity-based selection, the buffer containing PBS, EDTA, and human serum albumin.The wash was carried out within a sterile, single-use disposable kit sold by Biosafe SA for use in regenerative medicine, which included a centrifugal processing chamber (A-200F). The transfer pack containing the cells and a bag containing the buffer were sterilely connected to the kit, which was placed in association with a Sepax® 2 processing unit.
……….略

Beads were mixed in selection buffer described above, which then was sterilely connected to the apparatus. A program was run on the Sepax® 2 unit which caused the bead mixture and selection buffer to be drawn into the chamber with the washed cells, and the contents of the chamber to be mixed for 30 minutes.

Juno Therapeutics, Inc., Production of engineered cells for adoptive cell therapy, WO/2018/106732, 2018-06-14

無縫接軌-讓研究結果更快導入市場

Sepax能夠為使用者降低細胞製程的作業難度、提昇生產效率,同時封閉式賦予更好的安全性,有助產品的推行。而這麼多代表性的細胞治療大廠不約而同都選擇使用Sepax細胞血球分離機於各自的製程中,實則證明Sepax的技術、適用性均受世界頂尖細胞療法公司首肯、認同。

總的來說,Sepax的優點可以歸納成以下7點:
多用途 – 使用具彈性。
高再現 – 減少人為介入,產出高一致性。
高擴充 – 隨時可依需求擴充軟體、產能。
高效率 – 依功能不同僅需執行30~120分鐘。
高安全 – 封閉式系統、一次性耗材。
小巧輕變 – 容易導入現行設施規劃。
合規 – GMP-compatible

參考資料:
[1].Novartis Ag, Methods of making chimeric antigen receptor -expressing cells, WO/2017/117112, 2017-07-06
[2].Lu,TangyingLilyet al, “A Rapid Cell Expansion Process for Production of Engineered Autologous CAR-T Cell Therapies.”, Human gene therapy methods vol. 27,6 (2016): 209-218. doi:10.1089/hgtb.2016.120
[3].Kite Pharma, Inc., Methods for producing autologous T cells useful to treat B cell Malignancies and other cancers and compositions thereof, WO/2015/120096A3, 2015-11-12
[4].Juno Therapeutics, Inc., Production of engineered cells for adoptive cell therapy, WO/2018/106732, 2018-06-14

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