Rapid high-performance liquid chromatographic quantification of recombinant human antithrombin III during production and purification

Büntemeyer H, Tebbe H, Lütkemeyer D, Lehmann J. Rapid high-performance liquid chromatographic quantification of recombinant human antithrombin III during production and purification. Journal of Chromatography, B: Biomedical Sciences and Applications. 1994;662(2):209-216.For monitoring of recombinant human antithrombin III during cell culture processes and subsequent purification steps a rapid method for quantitative determination was developed. The need for the introduction of this rapid method came from the limited availability of a quantitative enzyme-linked immunosorbent assay (ELISA) and the very time-consuming ELISA procedure. The developed method is based on reversed-phase high-performance liquid chromatography using a C 4 column. The separation by gradient elution using water and acetonitrile takes less than 20 min even when complex samples, such as serum containing cell culture samples, have to be analyzed. Automation and a high sample throughput are possible with this reliable method. If necessary, insulin, transferrin and albumin can also be quantified with minor changes of the elution profile

Re-use of spent cell culture medium in pilot scale and rapid preparative purification with membrane chromatography

Riese U, Lütkemeyer D, Heidemann R, Büntemeyer H, Lehmann J. Re-use of spent cell culture medium in pilot scale and rapid preparative purification with membrane chromatography. Journal of Biotechnology. 1994;34(3):247-257.Based on experiments in bench scale, a recycling of spent cell culture medium was performed in a 100-l pilot scale bioreactor. The cell cultivation has been done as a repeated batch procedure after the initial batch in the following four repeated batches spent medium from the previous batch was partially re-used. After microfiltration and ultrafiltration a part of the filtrate was mixed with a concentrate of amino acids and glucose, sterile filtered and subsequently filled back into the bioreactor. Up to 65% of the harvested cell- and product-free spent medium was re-used in each repeated batch. This procedure results in a saving of pure and waste water volume and saving of supplemented proteins as transferrin, insulin and lipoproteins and, therefore, also in a reduction of the production costs. A strongly acidic membrane ion exchanger was evaluated for the ability to purify the monoclonal antibodies from the pilot scale cultivation. Within minutes, gram quantities of product could be purified in a high flux system, especially developed for this purpose, achieving purities of 80%. The capacity of the acidic membrane ion exchanger was found in former investigations to be 1 mg cm -2 with recoveries up to 96%. Final purification was carried out by gel column filtration

A method for metabolomic sampling of suspended animal cells using fast filtration

Volmer M, Gettmann J, Scholz S, Büntemeyer H, Noll T. A method for metabolomic sampling of suspended animal cells using fast filtration. BMC Proceedings. 2011;5(8):95

Proteomic and metabolomic characterization of CHO DP-12 cell lines with different high passage histories

Beckmann T, Thüte T, Heinrich C, Büntemeyer H, Noll T. Proteomic and metabolomic characterization of DP-12 cell lines with different high passage histories. In: BMC Proceedings. BMC Proceedings. Vol 5. BioMed Central; 2011

Ein Zellkulturfermenter mit integriertem Membransystem für die homogene Kulturführung

Büntemeyer H, Bödeker BGD, Lehmann J. Ein Zellkulturfermenter mit integriertem Membransystem für die homogene Kulturführung. Chemie - Ingenieur - Technik. 1990;62(5):393-395

Utilization of multifrequency permittivity measurements in addition to biomass monitoring

Heinrich C, Beckmann T, Büntemeyer H, Noll T. Utilization of multifrequency permittivity measurements in addition to biomass monitoring. BMC Proceedings. 2011;5(Suppl 8)

Utilization of multifrequency permittivity measurements in addition to biomass monitoring

Heinrich C, Beckmann T, Büntemeyer H, Noll T. Utilization of multifrequency permittivity measurements in addition to biomass monitoring. BMC Proceedings. 2011;5(Suppl 8)

Characterization of the human AGE1.HN cell line: a systems biology approach

Scholz S, Luebbecke M, Rath A, et al. Characterization of the human AGE1.HN cell line: a systems biology approach. BMC Proceedings. 2011;5(8)

The avian cell line AGE1.CR.pIX characterized by metabolic flux analysis

Lohr V, Haedicke O, Genzel Y, et al. The avian cell line AGE1.CR.pIX characterized by metabolic flux analysis. BMC Biotechnology. 2014;14(1): 72.Background: In human vaccine manufacturing some pathogens such as Modified Vaccinia Virus Ankara, measles, mumps virus as well as influenza viruses are still produced on primary material derived from embryonated chicken eggs. Processes depending on primary cell culture, however, are difficult to adapt to modern vaccine production. Therefore, we derived previously a continuous suspension cell line, AGE1.CR.pIX, from muscovy duck and established chemically-defined media for virus propagation. Results: To better understand vaccine production processes, we developed a stoichiometric model of the central metabolism of AGE1.CR.pIX cells and applied flux variability and metabolic flux analysis. Results were compared to literature dealing with mammalian and insect cell culture metabolism focusing on the question whether cultured avian cells differ in metabolism. Qualitatively, the observed flux distribution of this avian cell line was similar to distributions found for mammalian cell lines (e.g. CHO, MDCK cells). In particular, glucose was catabolized inefficiently and glycolysis and TCA cycle seem to be only weakly connected. Conclusions: A distinguishing feature of the avian cell line is that glutaminolysis plays only a minor role in energy generation and production of precursors, resulting in low extracellular ammonia concentrations. This metabolic flux study is the first for a continuous avian cell line. It provides a basis for further metabolic analyses to exploit the biotechnological potential of avian and vertebrate cell lines and to develop specific optimized cell culture processes, e.g. vaccine production processes