scholarly journals The Comparison of Chemiluminescent- and Colorimetric-detection Based ELISA for Chinese Hamster Ovary Host Cell Proteins Quantification in Biotherapeutics

2013 ◽  
Vol 03 (03) ◽  
Author(s):  
Fengqiang Wang Dennis Driscoll
2018 ◽  
Vol 13 (10) ◽  
pp. 1800111 ◽  
Author(s):  
Thomas Amann ◽  
Anders Holmgaard Hansen ◽  
Stefan Kol ◽  
Gyun Min Lee ◽  
Mikael Rørdam Andersen ◽  
...  

BioTechniques ◽  
2020 ◽  
Vol 69 (3) ◽  
pp. 186-192
Author(s):  
Kathleen Van Manen-Brush ◽  
Jacob Zeitler ◽  
John R White ◽  
Paul Younge ◽  
Samantha Willis ◽  
...  

Chinese hamster ovary (CHO) cells are a mammalian cell line used in the production of therapeutic proteins. Host cell proteins (HCPs) are process-related impurities that are derived from the host cell expression system. During biopharmaceutical drug development, removal of HCPs is required. Enzyme-linked immunosorbent assay (ELISA) is a common technique to quantitate HCPs, but is a labor-intensive process that takes up to 7 h. Ella® is an automated instrument that utilizes microfluidics and glass nanoreactors to quantitate HCPs in 75 min using similar ELISA reagents. The antibodies and antigens are captured on three distinct glass nanoreactors, resulting in sensitive reproducible data. Our results indicate that Ella quantitates CHO HCPs with precision, accuracy, sensitivity and trends comparable with our traditional CHO HCP ELISA.


2019 ◽  
Vol 20 (7) ◽  
pp. 1729 ◽  
Author(s):  
R. Lavoie ◽  
Alice di Fazio ◽  
R. Blackburn ◽  
Michael Goshe ◽  
Ruben Carbonell ◽  
...  

The growing integration of quality-by-design (QbD) concepts in biomanufacturing calls for a detailed and quantitative knowledge of the profile of impurities and their impact on the product safety and efficacy. Particularly valuable is the determination of the residual level of host cell proteins (HCPs) secreted, together with the product of interest, by the recombinant cells utilized for production. Though often referred to as a single impurity, HCPs comprise a variety of species with diverse abundance, size, function, and composition. The clearance of these impurities is a complex issue due to their cell line to cell line, product-to-product, and batch-to-batch variations. Improvements in HCP monitoring through proteomic-based methods have led to identification of a subset of “problematic” HCPs that are particularly challenging to remove, both at the product capture and product polishing steps, and compromise product stability and safety even at trace concentrations. This paper describes the development of synthetic peptide ligands capable of capturing a broad spectrum of Chinese hamster ovary (CHO) HCPs with a combination of peptide species that allow for advanced mixed-mode binding. Solid phase peptide libraries were screened for identification and characterization of peptides that capture CHO HCPs while showing minimal binding of human IgG, utilized here as a model product. Tetrameric and hexameric ligands featuring either multipolar or hydrophobic/positive amino acid compositions were found to be the most effective. Tetrameric multipolar ligands exhibited the highest targeted binding ratio (ratio of HCP clearance over IgG loss), more than double that of commercial mixed-mode and anion exchange resins utilized by industry for IgG polishing. All peptide resins tested showed preferential binding to HCPs compared to IgG, indicating potential uses in flow-through mode or weak-partitioning-mode chromatography.


2019 ◽  
Vol 117 (2) ◽  
pp. 438-452 ◽  
Author(s):  
R. Ashton Lavoie ◽  
Alice Fazio ◽  
Taufika Islam Williams ◽  
Ruben Carbonell ◽  
Stefano Menegatti

2013 ◽  
Vol 9 (1) ◽  
pp. 87-99 ◽  
Author(s):  
Kristin N. Valente ◽  
Amy K. Schaefer ◽  
Hannah R. Kempton ◽  
Abraham M. Lenhoff ◽  
Kelvin H. Lee

Mutagenesis ◽  
1997 ◽  
Vol 12 (4) ◽  
pp. 277-283 ◽  
Author(s):  
Rhonda L. Rolig ◽  
Susan K. Layher ◽  
Barbara Santi ◽  
Gerald M. Adair ◽  
Feng Gu ◽  
...  

2004 ◽  
Vol 24 (6) ◽  
pp. 559-576 ◽  
Author(s):  
Lili Liu ◽  
Andrew J. Rainbow

We have used a non-replicating recombinant adenovirus, Ad5MCMVlacZ, which expresses the β-galactosidase reporter gene, to examine both constitutive and inducible repair of UV-damaged DNA in repair proficient CHO-AA8 Chinese hamster ovary cells and in mutant CHO-UV61 cells which are deficient in the transcription-coupled repair (TCR) pathway of nucleotide excision repair. Host cell reactivation (HCR) of β-galactosidase activity for UV-irradiated Ad5MCMVlacZ was significantly reduced in non-irradiated CHO-UV61 cells compared to that in non-irradiated CHO-AA8 cells suggesting that repair in the transcribed strand of the UV-damaged reporter gene in untreated cells utilizes TCR. Prior UV-irradiation of cells with low UV fluences resulted in a transient enhancement of HCR for expression of the UV-damaged reporter gene in CHO-AA8 cells but not in TCR deficient CHO-UV61 cells. These results suggest the presence of an inducible DNA pathway in CHO cells that results from an enhancement of TCR or a mechanism that involves the TCR pathway.


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