Monitoring Glycosylation Profile and Protein Titer in Cell Culture Samples Using ZipChip CE-MS

Wang, Yan; Peng, Feng; Sosic, Zoran; Zang, Li

doi:10.4172/2155-9872.1000359

Cited by 26 publications

(31 citation statements)

References 16 publications

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“…The ZipChip system was operated according to manufacturer protocol for high resolution separation as previously described 15 . Briefly, the device was positioned on the inlet of a Q Exactive HF (Thermo).…”

Section: Methodsmentioning

confidence: 99%

Assessing a commercial capillary electrophoresis interface (ZipChip) for shotgun proteomic applications

Rinas

Jenkins

Orsburn

2019

Preprint

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Capillary electrophoresis coupled electrospray ionization mass spectrometry (CE-MS) has long existed as a theoretical alternative to liquid chromatography coupled mass spectrometry (LC-MS). Until recently, however, the coupling of these technologies has occupied only a small niche within specific applications. A recent innovation in CE-MS is the ZipChip interface system from 908 devices that was pioneered by the Ramsay lab at NC State. This newly available source offers advantages over previous CE-MS interfaces including both relative ease of use and direct compatibility to thousands of mass spectrometers currently in use throughout the world with no hardware alterations. The ZipChip CE-MS has been demonstrated in recent studies to provide high resolution and rapid separations for the analysis of intact proteins, glycoproteins and glycosylated peptides, with more applications likely on the way. In this study we assess the capabilities of the ZipChip system in the context of high throughput global shotgun proteomics experiments. We find that on a high field Orbitrap system we can repeatedly identify as many as 800 unique protein groups in an experiment using a run time of 12 minutes. We find the ZipChip CE-MS system to be widely applicable for both data dependent and data independent acquisition experiments as well as targeted experiments. We conclude that the ZipChip is an attractive alternative solution to traditional nanoflow ESI-MS/MS for the analysis of the genomes of single celled organisms and for offline fractionation of eukaryotic proteomes.

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Section: Methodsmentioning

confidence: 99%

Assessing a commercial capillary electrophoresis interface (ZipChip) for shotgun proteomic applications

Rinas

Jenkins

Orsburn

2019

Preprint

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“…[2][3][4] The main objective is to real time monitor the concentrations of some process parameters, such as viable cell, nutrient, and metabolite concentrations, whose variability may have an impact on mAb quality attributes. [10][11][12] Moreover, delays due to off-line analysis may also compromise real time control of the process. [6][7][8] Therefore, its control is essential to ensure efficacy of the product and safety for patients.…”

Section: Introductionmentioning

confidence: 99%

“…9 Even in advanced production processes supporting the PAT initiative, glycosylationmonitoring still requires some sample handling through the semiautomatic off-line analysis. [10][11][12] Moreover, delays due to off-line analysis may also compromise real time control of the process. A new challenging objective for PAT is thus to control mAb glycosylation as well as cell metabolism using online spectroscopy.…”

Section: Introductionmentioning

confidence: 99%

Interest of locally weighted regression to overcome nonlinear effects during in situ NIR monitoring of CHO cell culture parameters and antibody glycosylation

Zavala-Ortiz

Ebel

et al. 2019

Biotechnology Progress

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Animal cell culture processes have become the standard platform to produce therapeutic proteins such as recombinant monoclonal antibodies (mAb). Since the mAb quality could be subject to significant changes depending on manufacturing process conditions, real time monitoring and control systems are required to ensure mAb specifications mainly glycosylation and patient safety. Up to now, real time monitoring glycosylation of proteins has received scarce attention. In this article, the use of near infrared (NIR) to monitor mAb glycosylation has been reported for the first time.Whereas monitoring models are mainly constructed using linear partial least squares regressions (PLSR), evidences presented in this study indicate nonlinearity relationship between in situ captured spectra and compound concentrations, compromising the PLSR performances. A novel and simple approach was proposed to fit nonlinearity using the locally weighted regression (LWR). The LWR models were found to be more appropriate for handling information contained in spectra so that real time monitoring of cultures were accurately performed. Moreover, for the first time, the LWR calibration models allowed mAb glycosylation to be monitored, in a real time manner, by using in situ NIR spectroscopy. These results represent a further step toward developing active-control feedback of animal cell processes, particularly for ensuring properties of biologics.in situ near infrared spectroscopy, locally weighted regression-LWR, monoclonal antibody-mAb glycosylation, partial least squares regression-PLSR, real time monitoring

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“…and Zang et al. . In utilization of the HR variety chip, intact antibodies have been analyzed as well as viral capsid proteins by Chen et al.…”

Section: Introductionmentioning

confidence: 99%

Microfluidic capillary zone electrophoresis mass spectrometry analysis of alkaloids in Lobelia cardinalis transgenic and mutant plant cell cultures

et al. 2019

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Application of a microfluidic CE* device for CZE‐MS allows for fast, rapid, and in‐depth analysis of large sample sets. This microfluidic CZE‐MS device, the 908 Devices ZipChip, involves minimal sample preparation and is ideal for small cation analytes, such as alkaloids. Here, we evaluated the microfluidic device for the analysis of alkaloids from Lobelia cardinalis hairy root cultures. Extracts from wild‐type, transgenic, and selected mutant plant cultures were analyzed and data batch processed using the mass spectral processing software MZmine2 and the statistical software Prism 8. In total 139 features were detected as baseline resolved peaks via the MZmine2 software optimized for the electrophoretic separations. Statistically significant differences in the relative abundance of the primary alkaloid lobinaline (C27H34N2), along with several putative “lobinaline‐like” molecules were observed utilizing this approach. Additionally, a method for performing both targeted and untargeted MS/MS experiments using the microfluidic device was developed and evaluated. Coupling data‐processing software with CZE‐MS data acquisition has enabled comprehensive metabolomic profiles from plant cell cultures to be constructed within a single working day.

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Monitoring Glycosylation Profile and Protein Titer in Cell Culture Samples Using ZipChip CE-MS

Cited by 26 publications

References 16 publications

Assessing a commercial capillary electrophoresis interface (ZipChip) for shotgun proteomic applications

Assessing a commercial capillary electrophoresis interface (ZipChip) for shotgun proteomic applications

Interest of locally weighted regression to overcome nonlinear effects during in situ NIR monitoring of CHO cell culture parameters and antibody glycosylation

Microfluidic capillary zone electrophoresis mass spectrometry analysis of alkaloids in Lobelia cardinalis transgenic and mutant plant cell cultures

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