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  • 發布時間:2020-04-27 22:24 原文鏈接: offtheShelfT細胞治療技術

    自體CAR-T細胞過繼轉移可誘導復發/難治性血液惡性腫瘤患者的持續緩解,但是同時也存在一系列問題:1. 重癥癌癥病人,可能無法獲取自體T細胞進行CAR-T生產 2. 自體CAR-T細胞的異質性可能帶來不可預測的臨床反應。

    Precision BioScience通過其ZLARCUS基因編輯技術平臺,使用健康人T細胞建立通用CAR-T細胞,獲得了生物制藥商Baxalta17億美金投資。通過基因編輯去除細胞原有TCR,避免內源TCR引起的移植排斥反應,通過基因編輯,改變免疫耐受,提高CAR-T到腫瘤局部的能力,且具有保證批間一致性的優勢。

    Precision BioScience 在2018年三月份Best Practice & Research Clinical Haematology上發表了其off-the-Shelf T細胞治療技術體系Off the shelf T cell therapies for hematologic malignancies。

    Off the shelf T cell技術流程:文章系統介紹了Off the shelf T cell技術

    1. Gene editing for production of allogeneic CAR-T cells from unrelated healthy donor cells

    2. Allogeneic CAR-T cells with targeted CAR insertion by homologous recombination

    3. Additional gene edits to reduce immunogenicity and resist suppression within the tumor microenvironment

    4. Allogeneic CAR-T cell manufacturing

    5. Allogeneic CAR-T cell analytical methods

    5.1 Viability and cell count

    5.2 Cellular phenotype

    5.3 Functional analysis

    Traditionally, functional response of any T cell to antigenic challenges is estimated by cytokine production, cytotoxic activity, and T cell proliferation. Interferon gamma is a classical target to analyze T cell potency and is utilized as indicative of the biological functionality of manufactured T cell products. Conventional ELISA is a time-consuming procedure which requires well trained personnel or additional expenses for the automatization of the entire procedure. The Microfluidic Simple Plex cartridge-based assay on the ELLA instrument (ProteinSimple, San Jose, CA) may be a viable alternative to traditional ELISA. The entire procedure is handsfree and take approximately 90 min for readout.

    在功能分析( Functional analysis)方面Precision Medicine特地提到了使用Ella全自動的微流體ELISA系統來替代傳統ELISA方法。

    Ella在細胞功能分析及細胞因子風暴檢測的三大優勢:

    1. 超敏:IL-6等可達fg/ml,為傳統ELISA 10-100倍

    2. 快速:從樣本處理到結構,90mins之內

    3.全自動:降低實驗間CV

    之前,Ella 系統已經被Novartis和Kite Pharma等細胞治療的公司用作細胞功能質量控制。


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