71 results found
Goey CH, Bell D, Kontoravdi C, 2018, Mild hypothermic culture conditions affect residual host cell protein composition post-Protein A chromatography., MAbs, Pages: 1-12
Host cell proteins (HCPs) are endogenous impurities, and their proteolytic and binding properties can compromise the integrity, and, hence, the stability and efficacy of recombinant therapeutic proteins such as monoclonal antibodies (mAbs). Nonetheless, purification of mAbs currently presents a challenge because they often co-elute with certain HCP species during the capture step of protein A affinity chromatography. A Quality-by-Design (QbD) strategy to overcome this challenge involves identifying residual HCPs and tracing their source to the harvested cell culture fluid (HCCF) and the corresponding cell culture operating parameters. Then, problematic HCPs in HCCF may be reduced by cell engineering or culture process optimization. Here, we present experimental results linking cell culture temperature and post-protein A residual HCP profile. We had previously reported that Chinese hamster ovary cell cultures conducted at standard physiological temperature and with a shift to mild hypothermia on day 5 produced HCCF of comparable product titer and HCP concentration, but with considerably different HCP composition. In this study, we show that differences in HCP variety at harvest cascaded to downstream purification where different residual HCPs were present in the two sets of samples post-protein A purification. To detect low-abundant residual HCPs, we designed a looping liquid chromatography-mass spectrometry method with continuous expansion of a preferred, exclude, and targeted peptide list. Mild hypothermic cultures produced 20% more residual HCP species, especially cell membrane proteins, distinct from the control. Critically, we identified that half of the potentially immunogenic residual HCP species were different between the two sets of samples.
Sou SN, Lee K, Nayyar K, et al., 2018, Exploring cellular behavior under transient gene expression and its impact on mAb productivity and Fc-glycosylation, BIOTECHNOLOGY AND BIOENGINEERING, Vol: 115, Pages: 512-518, ISSN: 0006-3592
Alhuthali S, Fadda S, Goey CH, et al., 2017, Multi-stage population balance model to understand the dynamics of fed-batch CHO cell culture, Pages: 2821-2826, ISSN: 1570-7946
© 2017 Elsevier B.V. Monoclonal antibodies are crucial in the treatment of a broad spectrum of critical illnesses such as cancer and autoimmune diseases. These biopharmaceuticals are industrially produced in fed-batch animal cell culture bioreactors to achieve high cell density and product titre. At the end of the culture, the valuable product is selectively separated from a wide range of impurities also present in the supernatant, most of which are released by the host cells. A better insight into the dynamics of host cell impurity accumulation would facilitate optimisation and control of process performance and product quality. A multi-stage population balance model was developed to simulate cell growth, viability and the lysis of dead cells in an effort to trace the profile of a particular class of host cell impurities known as host cell proteins (HCPs). The model was informed by data from antibody-producing Chinese hamster ovary (CHO) cell cultures. The simulation results support that more than 20% of dead cells lyse in the interval between consecutive cell counts, releasing a variety of HCPs into the supernatant. The model also calculates that approximately 20% of the total HCP amount found extracellularly are likely to be secreted by viable cells during culture. The proposed model can be used in process development to gain estimates of HCP concentration under different process conditions, such as different feeding strategies, culture temperature and harvest time. These conditions can also be optimised in silico to decrease the level of cell-derived impurities.
Cardenas-Fernandez M, Bawn M, Hamley-Bennett C, et al., 2017, An integrated biorefinery concept for conversion of sugar beet pulp into value-added chemicals and pharmaceutical intermediates, FARADAY DISCUSSIONS, Vol: 202, Pages: 415-431, ISSN: 1359-6640
Goers L, Ainsworth C, Goey CH, et al., 2017, Whole-cell Escherichia coli lactate biosensor for monitoring mammalian cell cultures during biopharmaceutical production, BIOTECHNOLOGY AND BIOENGINEERING, Vol: 114, Pages: 1290-1300, ISSN: 0006-3592
Cover Legend The cover image, by Lisa Goers et al., is based on the Article Whole-cell Escherichia coli lactate biosensor for monitoring mammalian cell cultures during biopharmaceutical production, DOI: 10.1002/bit.26254.
Goey CH, Tsang JMH, Bell D, et al., 2017, Cascading effect in bioprocessing-The impact of mild hypothermia on CHO cell behavior and host cell protein composition, BIOTECHNOLOGY AND BIOENGINEERING, Vol: 114, Pages: 2771-2781, ISSN: 0006-3592
Sou SN, Jedrzejewski PM, Lee K, et al., 2017, Model-based investigation of intracellular processes determining antibody Fc-glycosylation under mild hypothermia, BIOTECHNOLOGY AND BIOENGINEERING, Vol: 114, Pages: 1570-1582, ISSN: 0006-3592
Klymenko OV, Shah N, Kontoravdi C, et al., 2016, Designing an Artificial Golgi Reactor to Achieve Targeted Glycosylation of Monoclonal Antibodies, AICHE JOURNAL, Vol: 62, Pages: 2959-2973, ISSN: 0001-1541
Niu H, Shah N, Kontoravdi C, 2016, Modelling of amorphous cellulose depolymerisation by cellulases, parametric studies and optimisation, BIOCHEMICAL ENGINEERING JOURNAL, Vol: 105, Pages: 455-472, ISSN: 1369-703X
del Val IJ, Polizzi KM, Kontoravdi C, 2016, A theoretical estimate for nucleotide sugar demand towards Chinese Hamster Ovary cellular glycosylation, SCIENTIFIC REPORTS, Vol: 6, ISSN: 2045-2322
Fan Y, Del Val IJ, Mueller C, et al., 2015, Amino Acid and Glucose Metabolism in Fed-Batch CHO Cell Culture Affects Antibody Production and Glycosylation, BIOTECHNOLOGY AND BIOENGINEERING, Vol: 112, Pages: 521-535, ISSN: 0006-3592
Fan Y, Del Val IJ, Muller C, et al., 2015, A multi-pronged investigation into the effect of glucose starvation and culture duration on fed-batch CHO cell culture, BIOTECHNOLOGY AND BIOENGINEERING, Vol: 112, Pages: 2172-2184, ISSN: 0006-3592
Niv H, Leak D, Shah N, et al., 2015, Metabolic characterization and modeling of fermentation process of an engineered Geobacillus thermoglucosidasius strain for bioethanol production with gas stripping, CHEMICAL ENGINEERING SCIENCE, Vol: 122, Pages: 138-149, ISSN: 0009-2509
Polizzi KM, Kontoravdi C, 2015, Genetically-encoded biosensors for monitoring cellular stress in bioprocessing, CURRENT OPINION IN BIOTECHNOLOGY, Vol: 31, Pages: 50-56, ISSN: 0958-1669
Sou SN, Sellick C, Lee K, et al., 2015, How does mild hypothermia affect monoclonal antibody glycosylation?, Biotechnology and Bioengineering, Vol: 112, Pages: 1165-1176, ISSN: 1097-0290
Jedrzejewski PM, del Val IJ, Constantinou A, et al., 2014, Towards Controlling the Glycoform: A Model Framework Linking Extracellular Metabolites to Antibody Glycosylation, INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, Vol: 15, Pages: 4492-4522, ISSN: 1422-0067
Kyriakopoulos S, Kontoravdi C, 2014, A Framework for the Systematic Design of Fed-Batch Strategies in Mammalian Cell Culture, BIOTECHNOLOGY AND BIOENGINEERING, Vol: 111, Pages: 2466-2476, ISSN: 0006-3592
Kyriakopoulos S, Kontoravdi C, 2014, Insights on biomarkers from Chinese hamster ovary ‘omics’ studies, Pharmaceutical Bioprocessing, Vol: 2, Pages: 389-401, ISSN: 2048-9145
Todri E, Amenaghawon AN, del Val IJ, et al., 2014, Global sensitivity analysis and meta-modeling of an ethanol production process, CHEMICAL ENGINEERING SCIENCE, Vol: 114, Pages: 114-127, ISSN: 0009-2509
Chen N, Bennett MH, Kontoravdi C, 2013, Analysis of Chinese hamster ovary cell metabolism through a combined computational and experimental approach, Cytotechnology, Vol: Online First
Jedrzejewski PM, Jimenez del Val I, Polizzi KM, et al., 2013, Applying quality by design to glycoprotein therapeutics: experimental and computational efforts of process control, Pharmaceutical Bioprocessing, Vol: 1, Pages: 51-69
Jimenez Del Val I, Constantinou A, Dell A, et al., 2013, An integrated mechanistic model for nucleotide sugar metabolism and monoclonal antibody glycosylation, Pages: 658-659
Jimenez del Val I, Constantinou A, Dell A, et al., 2013, A quantitative and mechanistic model for monoclonal antibody glycosylation as a function of nutrient availability during cell culture, 23rd European Society for Animal Cell Technology (ESACT) Meeting: Better Cells for Better Health
Kontoravdi C, 2013, Systematic methodology for the development of mathematical models for biological processes., Methods Mol Biol, Vol: 1073, Pages: 177-190
Synthetic biology gives researchers the opportunity to rationally (re-)design cellular activities to achieve a desired function. The design of networks of pathways towards accomplishing this calls for the application of engineering principles, often using model-based tools. Success heavily depends on model reliability. Herein, we present a systematic methodology for developing predictive models comprising model formulation considerations, global sensitivity analysis, model reduction (for highly complex models or where experimental data are limited), optimal experimental design for parameter estimation, and predictive capability checking. Its efficacy and validity are demonstrated using an example from bioprocessing. This approach systematizes the process of developing reliable mathematical models at a minimum experimental cost, enabling in silico simulation and optimization.
Kontoravdi C, Samsatli NJ, Shah N, 2013, Development and design of bio-pharmaceutical processes, CURRENT OPINION IN CHEMICAL ENGINEERING, Vol: 2, Pages: 435-441, ISSN: 2211-3398
Kyriakopoulos S, Polizzi KM, Kontoravdi C, 2013, Comparative analysis of amino acid metabolism and transport in CHO variants with different levels of productivity, JOURNAL OF BIOTECHNOLOGY, Vol: 168, Pages: 543-551, ISSN: 0168-1656
Kyriakopoulos S, Polizzi KM, Kontoravdi C, 2013, Dynamic profiling of amino acid transport and metabolism in Chinese hamster ovary cell culture., Pages: P97-P97, ISSN: 1753-6561
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