Integrated Clarification and Purification of Monoclonal Antibodies by Membrane Based Separation of Aqueous Two-Phase Systems

被引:14
作者
Kruse, Thomas [1 ,2 ]
Schmidt, Axel [1 ]
Kampmann, Markus [2 ]
Strube, Jochen [1 ]
机构
[1] Tech Univ Clausthal, Inst Separat & Proc Technol, Leibnizstr 15, D-38678 Clausthal Zellerfeld, Germany
[2] Sartorius Sted Biotech GmbH, August Spindler Str 11, D-37079 Gottingen, Germany
关键词
aqueous two-phase extraction; phase separation; downstream; clarification; membrane; CENTRIFUGAL PARTITIONING CHROMATOGRAPHY; EXTRACTION; TECHNOLOGY; OPTIMIZATION; PERFORMANCE; PLATFORM; DESIGN;
D O I
10.3390/antib8030040
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Therapeutic monoclonal antibodies (mAb) are used for the treatment of numerous serious diseases, which have led to an increasing demand over the last decades. Increased cell density and mAb titer of the cultivation broth lead to great challenges for the subsequent clarification and capture operations in the downstream process. As an alternative approach to the conventional downstream process, a selective mAb extraction via an aqueous two-phase system (ATPS) directly from the cultivation broth of a mAb producing industrial relevant chinese hamster ovary (CHO) cell line was investigated. An efficient purification of the mAb was accomplished by the ATPS composition. The phase separation was realized by a newly developed membrane based phase separator. Moreover, a complete cell removal was integrated into this process by the used membrane. A selectivity between both phases was achieved by membrane modification. Yields up to 93% in the light phase and removal of process related impurities were obtained after aqueous two-phase extraction (ATPE). Phase separation performance as well as contact angles on the membrane were characterized for different ATPS. ATPE directly from the cultivation broth in combination with the new membrane based phase separation led to a mAb yield of 78% with a simultaneous reduction of deoxyribonucleic acid (DNA) and host cell protein (HCP) load.
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页数:17
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共 46 条
  • [1] Monoclonal antibodies in immune and inflammatory diseases
    Andreakos, E
    Taylor, PC
    Feldmann, M
    [J]. CURRENT OPINION IN BIOTECHNOLOGY, 2002, 13 (06) : 615 - 620
  • [2] ATHA DH, 1981, J BIOL CHEM, V256, P2108
  • [3] Integrated process for the purification of antibodies combining aqueous two-phase extraction, hydrophobic interaction chromatography and size-exclusion chromatography
    Azevedo, A. M.
    Rosa, P. A. J.
    Ferreira, I. F.
    Aires-Barros, M. R.
    [J]. JOURNAL OF CHROMATOGRAPHY A, 2008, 1213 (02) : 154 - 161
  • [4] Optimisation of aqueous two-phase extraction of human antibodies
    Azevedo, Ana M.
    Rosa, Paula A. J.
    Ferreira, I. Filipa
    Aires-Barros, M. Raquel
    [J]. JOURNAL OF BIOTECHNOLOGY, 2007, 132 (02) : 209 - 217
  • [5] Chromatography-free recovery of biopharmaceuticals through aqueous two-phase processing
    Azevedo, Ana M.
    Rosa, Paula A. J.
    Ferreira, I. Filipa
    Aires-Barros, M. Raquel
    [J]. TRENDS IN BIOTECHNOLOGY, 2009, 27 (04) : 240 - 247
  • [6] Belter P.A., 1988, Bioseparations: Downstream Processing for Biotechnology
  • [7] Experimental Study of Membrane Fouling during Crossflow Microfiltration of Yeast and Bacteria Suspensions: Towards an Analysis at the Microscopic Level
    Ben Hassan, Ines
    Ennouri, Monia
    Lafforgue, Christine
    Schmitz, Philippe
    Ayadi, Abdelmoneim
    [J]. MEMBRANES, 2013, 3 (02) : 44 - 68
  • [8] Continuous cell flocculation for recombinant antibody harvesting
    Burgstaller, Daniel
    Krepper, Walpurga
    Haas, Josselyn
    Maszelin, Marine
    Mohoric, Jure
    Pajnic, Katja
    Jungbauer, Alois
    Satzer, Peter
    [J]. JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY, 2018, 93 (07) : 1881 - 1890
  • [9] Recent advances in technology supporting biopharmaceutical production from mammalian cells
    Butler, M.
    Meneses-Acosta, A.
    [J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2012, 96 (04) : 885 - 894
  • [10] Cheryan M., 1990, Handbuch Ultrafiltration