Engineering selection stringency on expression vector for the production of recombinant human alpha1-antitrypsin using Chinese Hamster Ovary cells

Chin, Christine Lin; Chin, Hing Kah; Chin, Cara Sze Hui; Lai, Ethan Tingfeng; KongNg, Say

doi:10.1186/1753-6561-9-s9-p10

Cited by 3 publications

(4 citation statements)

References 22 publications

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“…Since ELISA is the most widely used assay for monitoring protein titer, it seems likely that a considerable number of reported recombinant protein titers is inaccurate. Recently, a r α1AT titer of 1.15 g/L was reported , which according to the authors is the highest titer of any recombinant protein in shake flasks batch culture of stable mammalian cells. A commercially available ELISA kit was used for determining protein titer and validation of the ELISA kit by an orthogonal assay was not described.…”

Section: Discussionmentioning

confidence: 99%

“…ra1AT can be produced in transgenic sheep [7], but an immune response to endogenous sheep a1AT in the purified product has been observed [8]. An attractive alterna-tive is to produce ra1AT in CHO or human cells and efforts have been undertaken to achieve this [4,[9][10][11][12][13][14]. N-glycosylation patterns of ra1AT produced from CHO and human cells have been shown to be similar but not identical to pla1AT [4,15].…”

Section: Introductionmentioning

confidence: 99%

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Case study on human α1‐antitrypsin: Recombinant protein titers obtained by commercial ELISA kits are inaccurate

Hansen

Kildegaard

Lee

et al. 2016

Biotechnology Journal

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Case study on human α1‐antitrypsin: Recombinant protein titers obtained by commercial ELISA kits are inaccurate

Hansen

Kildegaard

Lee

et al. 2016

Biotechnology Journal

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“…Recombinant AAT (rAAT) was produced in CHO-DG44 cells by the Animal Cell Technology (ACT) group at the Bioprocessing Technology Institute (BTI), as described previously (Chin, Chin, Chin, Lai, & Ng, 2015). 1 An ÄKTA Purifier 10 (Amersham Pharmacia Biotech, United Kingdom) was used to purify the culture supernatant of rAAT.…”

Section: Recombinant Aat Productionmentioning

confidence: 99%

Ion-Mobility Mass Spectrometry of Glycans and Glycopeptides Using Both Intact and Glycan Sequencing Approaches In Positive Ion Mode Facilitates Glycan Isomer and Glycan Motif Identification

Pallister

Choo²,

Walsh³

et al. 2020

Preprint

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High resolution glycan analysis has become an important part of biopharmaceutical API production and quality control. Liquid chromatography (LC) is now a well-established technique in this field but the resolution of similar isomeric glycan structures is still a challenge. Here we show that the addition of ion mobility spectroscopy (IMS) in a hyphenated LC-IMS-MS setting allows for the high resolution of N-glycan isomers during positive ion analysis. We have identified unique features in the IM chromatograms to help differentiate a range of isomeric N-glycans for both RFMS labelled glycans and glycopeptides.<br>

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“…The extracellular concentrations after feeding were computed based on the feed composition information. Specific growth rate, m, was computed for each experimental condition separately as the slope of the linear trend line obtained by plotting ln(C XR .V R ) against time (Chin et al, 2015;Clarke et al, 2011).…”

Section: Cell Line Cell Cultivation Sampling and Rate Estimationsmentioning

confidence: 99%

Segmented linear modeling of CHO fed‐batch culture and its application to large scale production

Yahia

Gourévitch

Malphettes

et al. 2016

Biotech & Bioengineering

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We describe a systematic approach to model CHO metabolism during biopharmaceutical production across a wide range of cell culture conditions. To this end, we applied the metabolic steady state concept. We analyzed and modeled the production rates of metabolites as a function of the specific growth rate. First, the total number of metabolic steady state phases and the location of the breakpoints were determined by recursive partitioning. For this, the smoothed derivative of the metabolic rates with respect to the growth rate were used followed by hierarchical clustering of the obtained partition. We then applied a piecewise regression to the metabolic rates with the previously determined number of phases. This allowed identifying the growth rates at which the cells underwent a metabolic shift. The resulting model with piecewise linear relationships between metabolic rates and the growth rate did well describe cellular metabolism in the fed‐batch cultures. Using the model structure and parameter values from a small‐scale cell culture (2 L) training dataset, it was possible to predict metabolic rates of new fed‐batch cultures just using the experimental specific growth rates. Such prediction was successful both at the laboratory scale with 2 L bioreactors but also at the production scale of 2000 L. This type of modeling provides a flexible framework to set a solid foundation for metabolic flux analysis and mechanistic type of modeling. Biotechnol. Bioeng. 2017;114: 785–797. © 2016 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.

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Engineering selection stringency on expression vector for the production of recombinant human alpha1-antitrypsin using Chinese Hamster Ovary cells

Cited by 3 publications

References 22 publications

Case study on human α1‐antitrypsin: Recombinant protein titers obtained by commercial ELISA kits are inaccurate

Case study on human α1‐antitrypsin: Recombinant protein titers obtained by commercial ELISA kits are inaccurate

Ion-Mobility Mass Spectrometry of Glycans and Glycopeptides Using Both Intact and Glycan Sequencing Approaches In Positive Ion Mode Facilitates Glycan Isomer and Glycan Motif Identification

Segmented linear modeling of CHO fed‐batch culture and its application to large scale production

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