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具有自动克隆筛选功能的细胞株开发解决方案

ClonePix® 2 哺乳动物细胞克隆筛选系统是一个全自动系统,可用于选择在抗体发现和细胞株开发中使用的优质细胞克隆。对杂交瘤细胞、CHO 细胞以及其他细胞类型成像并根据用户定义的参数进行筛选。全面集成孔板处理、条形码读取以及克隆挑取功能。保存包括图像在内的所有数据,以用于下游分析。该筛选系统增加了找到稀有优质细胞株的概率并显著节省了时间和人力。

  • 高效

    在更短时间内筛选更多的克隆

    ClonePix 2 筛选系统要比劳动密集的有限稀释法和 FACS 快 10 倍。我们先进的软件和集成式机械设备可实现每天 > 10,000 个克隆的筛选速度。

  • 选择

    挑选具有所需特性的细胞

    根据蛋白质产率、抗原特异性、细胞活性以及已标记重组蛋白的表达水平来轻松筛选并挑取克隆。

  • 准确度

    精准挑取克隆

    挑取精度 < 1mm。机械挑取可降低克隆干扰的风险。已挑取克隆的图像和数据一并保存。

ClonePix 2 高通量细胞克隆筛选系统

ClonePix 2

特点

  • 多种检测方法

    白光可检测并测量克隆形态、尺寸和间距。荧光可指示表达水平和/或特异性。多达 5 种荧光滤光片可供使用。

  • 保持无菌性

    标准配置含一系列灭菌功能和选项,包括给仪器内部消毒的紫外灯,以及挑针清洗和卤素灯干燥。

  • 集成储板系统

    包括源板和终板的两个堆板架,每个可容纳 10 块板。

  • 离散的克隆形成

    半固体培养基 CloneMediaTM 支持将单细胞培养为离散的克隆,并支持轻松加样处理。该培养基可实现密度更高的克隆筛选。

  • 不含动物成分的培养基和试剂

    化学成分限定且不含动物成分的 CloneMedia 培养基经优化可增加产率,且在与 CloneDetectTM 检测试剂一同使用时有助于显示所分泌的抗体。

  • 定制自动化选项*

    先进工作流程工程解决方案团队可自定义单克隆系统并提供单克隆性综合验证等附加服务。

*价格、交付时间和技术参数将根据双方商定的技术要求而有所不同。可能会根据解决方案要求调整标准性能。

ClonePix 2 哺乳动物细胞克隆筛选系统的应用

  • 细胞株开发

    针对重组蛋白的细胞株株开发

    细胞株开发是制备生物制药分子(如单克隆抗体)过程中的关键步骤。该过程通常从将编码目标蛋白的 DNA 转染至宿主细胞开始,目标 DNA 会随机或定向整合至宿主细胞基因组中。筛选数以千计的克隆以鉴别稀有的高产细胞是一个手动且耗时的过程。

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    细胞表面表达筛选

    细胞表面表达筛选

    许多在细胞表面表达的蛋白都是用于发现和开发生物医药的靶标。例如,G 蛋白偶联受体 (GPCR) 是最大的细胞表面蛋白类别,并且是大约 40% 现有药物的靶标。从转染后的细胞池中发现和挑选细胞表面 GPCR 表达增加的高价值克隆可能极具挑战性。ClonePix 2 系统是可筛选大量细胞的一种自动化方法,它增加了发现稀有高亲和力杂交瘤细胞或高产细胞的概率。

  • 克隆产量筛选和滴度

    克隆产量筛选和滴度

    识别优质克隆的一个重要步骤是测定单细胞来源克隆的表达量。使用传统方法筛选产率费力且耗时,通常包含多个步骤,包括从有限稀释中分离单细胞之后再用 ELISA 进行滴度评估。ClonePix 2 系统将表型选择、单细胞分离和产率筛选合为一个步骤,因此可明显缩短筛选时间并增加候选物数量。

    杂交瘤细胞筛选

    杂交瘤细胞筛选

    抗体开发通常是指筛选和识别能靶向特定表位以实现疾病诊治的单克隆抗体 (mAb)。产生单克隆抗体的一种常用方法是将有丝分裂前的癌症细胞与脾脏中有丝分裂后和表达终端抗体的 B 细胞融合。产生的融合细胞称为杂交瘤细胞,它拥有产生单克隆抗体的优势,同时还可分裂再生。使用 ClonePix 2 系统可自动筛选杂交瘤细胞的结合特异性或产率。

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  • 单克隆抗体发现

    抗原特异性筛选

    抗体开发通常是指筛选和识别能靶向抗原分子以实现疾病诊治的特异性抗体。抗体的特异性取决于其结合表位(抗原分子上的独特区域)的能力。治疗性抗体通常为单细胞源性单克隆抗体,可靶向抗原上的独特表位区域。ClonePix 2 系统可从异源细胞群中自动筛选和快速检测抗原特异性克隆。

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    单克隆性

    细胞株开发和单克隆性验证是生产生物制药分子(如单克隆抗体)的过程中的关键步骤。在分离稳定表达目标蛋白的单个活细胞后,可建立细胞株。此过程中的一个关键结点是记录单克隆性证据。单克隆性证据通常都是图像形式,据此可生成单细胞图像并将其随附在监管文件中。

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ClonePix 2 哺乳动物细胞克隆筛选系统的技术参数和可选配置

ClonePix 2 哺乳动物细胞克隆筛选系统的资源

报告内容
视频和网络研讨会
开发和建立稳定细胞系

稳定的细胞系开发工作流程

杂交瘤工作流程

杂交瘤工作流程

分泌非单克隆抗体蛋白的哺乳动物细胞工作流程

选择高产分泌非单克隆抗体蛋白的哺乳动物细胞工作流程 - SynBioBeta 闪电演讲

迅速识别中和抗体

优化的工作流程可用于迅速区分中和抗体与病毒颗粒

G 蛋白偶联受体细胞系选择

使用 ClonePix 2 来识别和选择 GPCR 细胞系

ClonePix 2 高通量细胞克隆筛选系统

ClonePix 2

  • Citation
    Dated: Dec 01, 2017
    Publication Name: Journal of Immunological Methods

    Sequential screening by ClonePix FL and intracellular staining facilitate isolation of high producer cell lines for monoclonal antibody manufacturing

    Screening and characterization of cell lines for stable production are critical tasks in identifying suitable recombinant cell lines for the manufacture of protein therapeutics. To aid this essential function we have developed a methodology for the selection of antibody expressing cells using fluorescence based ClonePix FL colony isolation and… View more

    Screening and characterization of cell lines for stable production are critical tasks in identifying suitable recombinant cell lines for the manufacture of protein therapeutics. To aid this essential function we have developed a methodology for the selection of antibody expressing cells using fluorescence based ClonePix FL colony isolation and flow cytometry analysis following intracellular staining for immunoglobulin G (IgG). Our data show that characterization of cells by flow cytometry early in the clone selection process enables the identification of cell lines with the potential for high productivity and helps to eliminate unstable cell lines. We further demonstrate a correlation between specific productivity (qP) and intracellular heavy chain (HC) content with final productivity. The unique combination of screening using ClonePix FL and the flow cytometry approaches facilitated more efficient isolation of clonal cell lines with high productivity within a 15 week timeline and which can be applied across NS0 and CHO host platforms. Furthermore, in this study we describe the critical parameters for the ClonePix FL colony based selection and the associated calculations to provide an assessment of the probability of monoclonality of the resulting cell lines.

    Contributors: Gargi Roya, Guillermo Miro-Quesadab, Li Zhuanga, Tom Martina, Jie Zhub, Herren Wua, Marcello Marellia, Michael A.Bowena  
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  • Citation
    Dated: Dec 14, 2015
    Publication Name: 24th European Society for Animal Cell Technology (ESACT) Meeting: C2P2: Cells, Culture, Patients, Products

    CHO-DHFR cell line development platform: Application of Clonepix and Automated Mini Bioreactor (AMBR) technologies to meet accelerated timelines

    The Holy Grail sought by all Bioprocess Cell Line Development (CLD) groups is achieving high yields from easily-cultured, robustly-growing cells in timelines measured in weeks rather than months. As the first bottleneck in process development, CLD must first birth its product for upstream and downstream groups to initiate their own reproductive… View more

    The Holy Grail sought by all Bioprocess Cell Line Development (CLD) groups is achieving high yields from easily-cultured, robustly-growing cells in timelines measured in weeks rather than months. As the first bottleneck in process development, CLD must first birth its product for upstream and downstream groups to initiate their own reproductive cycles. To facilitate shortened CLD timelines, scientists have turned to new technologies and automation platforms. Emerging high-throughput instrumentation such as Clonepix and Automated MicroBioreactors (AMBR) have been enthusiastically integrated into stable cell line generation platforms; however, application of these methodologies among users is divergent.

    Contributors: Venkata R Mangalampalli, Dyane Wycuff, Mingzhong Chen, David Berlinger, Elizabeth H Scheideman, Amritha Menon, Guilia Fabozzi, Althaf Hussain & Richard M Schwartz  
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  • Citation
    Dated: May 25, 2014
    Publication Name: New Biotechnology

    High-throughput ClonePix FL analysis of mAb-expressing clones using the UCOE expression system

    Therapeutic recombinant monoclonal antibodies (mAbs) are commonly produced by high-expressing, clonal, mammalian cells. Creation of these clones for manufacturing remains heavily reliant on stringent selection and gene amplification, which in turn can lead to genetic instability, variable expression, product heterogeneity and prolonged development… View more

    Therapeutic recombinant monoclonal antibodies (mAbs) are commonly produced by high-expressing, clonal, mammalian cells. Creation of these clones for manufacturing remains heavily reliant on stringent selection and gene amplification, which in turn can lead to genetic instability, variable expression, product heterogeneity and prolonged development timelines. Inclusion of cis-acting ubiquitous chromatin opening elements (UCOE™) in mammalian expression vectors has been shown to improve productivity and facilitate high-level gene expression irrespective of the chromosomal integration site without lengthy gene amplification protocols. In this study we have used high-throughput robotic clone selection in combination with UCOE™ containing expression vectors to develop a rapid, streamlined approach for early-stage cell line development and isolation of high-expressing clones for mAb production using Chinese hamster ovary (CHO) cells. Our results demonstrate that it is possible to go from transfection to stable clones in only 4 weeks, while achieving specific productivities exceeding 20 pg/cell/day. Furthermore, we have used this approach to quickly screen several process-crucial parameters including IgG subtype, enhancer-promoter combination and UCOE™ length. The use of UCOE™-containing vectors in combination with automated robotic selection provides a rapid method for the selection of stable, high-expressing clones.

    Contributors: Jeff Jia Cheng Hou, Ben S. Hughes, Matthew Smede, Kar Man Leung, Kara Levine, Susan Rigby, Peter P. Gray, Trent P.Munro  
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