scholarly journals Accelerating Biomanufacturing by Modeling of Continuous Bioprocessing—Piloting Case Study of Monoclonal Antibody Manufacturing

Processes ◽  
2019 ◽  
Vol 7 (8) ◽  
pp. 495 ◽  
Author(s):  
Kornecki ◽  
Schmidt ◽  
Lohmann ◽  
Huter ◽  
Mestmäcker ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Final Analysis ◽  
Pilot Scale ◽  
Stable Operation ◽  
Two Phase ◽  
Ovary Cells ◽  
Cell Densities ◽  
Flow Filtration

An experimental feasibility study on continuous bioprocessing in pilot-scale of 1 L/day cell supernatant, that is, about 150 g/year product (monoclonal antibody) based on CHO (Chinese hamster ovary) cells for model validation is performed for about six weeks including preparation, start-up, batch, and continuous steady-state operation for at least two weeks stable operation as well as final analysis of purity and yield. A mean product concentration of around 0.4 g/L at cell densities of 25 × 106 cells/mL was achieved. After perfusion cultivation with alternating tangential flow filtration (ATF), an aqueous two-phase extraction (ATPE) followed by ultra-/diafiltration (UF/DF) towards a final integrated counter-current chromatography (iCCC) purification with an ion exchange (IEX) and a hydrophobic interaction (HIC) column prior to lyophilization were successfully operated. In accordance to prior studies, continuous operation is stable and feasible. Efforts of broadly-qualified operation personal as well as the need for an appropriate measurement and process control strategy is shown evidently.

scholarly journals Purification and Analytics of a Monoclonal Antibody from Chinese Hamster Ovary Cells Using an Automated Microbioreactor System

10.3791/58947 ◽  
2019 ◽  
Author(s):  
Sai Rashmika Velugula-Yellela ◽  
David N. Powers ◽  
Phillip Angart ◽  
Anneliese Faustino ◽  
Talia Faison ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Ovary Cells

Efficient enrichment of high-producing recombinant Chinese hamster ovary cells for monoclonal antibody by flow cytometry

2015 ◽  
Vol 120 (3) ◽  
pp. 340-346 ◽  
Author(s):  
Takeshi Okumura ◽  
Kenji Masuda ◽  
Kazuhiko Watanabe ◽  
Kenji Miyadai ◽  
Koichi Nonaka ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Flow Cytometry ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Ovary Cells

LC-MS-based metabolic characterization of high monoclonal antibody-producing Chinese hamster ovary cells

2012 ◽  
Vol 109 (12) ◽  
pp. 3103-3111 ◽  
Author(s):  
William Pooi Kat Chong ◽  
Shu Hui Thng ◽  
Ai Ping Hiu ◽  
Dong-Yup Lee ◽  
Eric Chun Yong Chan ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Ovary Cells

An integrated approach to analyze EG2-hFc monoclonal antibody N-glycosylation by MALDI-MS

2015 ◽  
Vol 93 (7) ◽  
pp. 754-763 ◽  
Author(s):  
Edward Bodnar ◽  
Thais Ferreira Nascimento ◽  
Lauren Girard ◽  
Emy Komatsu ◽  
Paul Lopez ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Chinese Hamster Ovary Cells ◽  
Protein A ◽  
Chinese Hamster ◽  
Integrated Approach ◽  
Maldi Ms ◽  
Ovary Cells ◽  
Pngase F ◽  
First Time

The characterization of the N-glycan portion of antibodies has been the subject of several studies involving mass spectrometry. In this article, a workflow is presented that starts with the expression of a monoclonal antibody (EG2-hFc) in Chinese hamster ovary cells and continues with Protein A purification of the antibody. Then the protocol continues with gel electrophoresis. Bands containing the heavy chain are cut and isolated from the gel followed by tryptic digestion to obtain peptides and glycopeptides. The enrichment of glycopeptides by C18 chromatography is described followed by characterization using positive and negative modes MALDI-MS and MS/MS. An exoglycosidase, beta-galactosidase, is used to verify anomericity of linkages in the glycan portion of glycopeptides. In the last step, glycans are detached from glycopeptides using PNGase F labelled with phehylhydrazine and characterized by MALDI-MS/MS. This workflow is reported for the first time for this particular antibody and presents a valuable approach for the analysis of N-glycans on most antibodies and glycoproteins.

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