Optimizing antibody production in batch hybridoma cell culture

McKinney, Katherine L.; Dilwith, Robert L.; Belfort, Georges

doi:10.1016/0168-1656(95)00030-t

Cited by 17 publications

(8 citation statements)

References 59 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These parameters included the heavy and light chain specific transcription and translation rates, which agrees with literature reports that stress the importance of the transcription (18,42,43) and translation (23,44) steps in the production mechanism of recombinant proteins (including antibodies). This is in contrast to the time required for intracellular transport of the antibody molecule from the ER to the golgi complex and subsequently the extracellular media, as reflected by the low sensitivity indices of t 1/2,ER and t 1/2,G .…”

Section: Discussionsupporting

confidence: 86%

Development and Analysis of a Mathematical Model for Antibody-Producing GS-NS0 Cells Under Normal and Hyperosmotic Culture Conditions

Varley

Mantalaris

2006

Biotechnol. Prog.

View full text Add to dashboard Cite

The GS-NS0 cell line is industrially important and is currently used for the large-scale production of several therapeutic monoclonal antibodies. A novel hybrid model, consisting of both unstructured and structured elements, has been developed to describe cell growth and death, metabolism, and antibody production in the GS-NS0 system under normal culture conditions. A comparison between the hybrid model and a large-scale single-cell model (SCM) describing detailed metabolic processes verified the predictive ability of the hybrid model (when compared with experimental data) and highlighted the practical difficulties involved in utilizing complex models. Global sensitivity analysis (GSA) on the hybrid model identified the specific transcription and translation rates of heavy and light immunoglobulin chains as parameters with the largest impact on the antibody production process. This information, together with the addition of a 24-h lag phase, resulted in the successful extension of the hybrid model to represent GS-NS0 system behavior under hyperosmotic culture conditions.

show abstract

Section: Discussionsupporting

confidence: 86%

Development and Analysis of a Mathematical Model for Antibody-Producing GS-NS0 Cells Under Normal and Hyperosmotic Culture Conditions

Varley

Mantalaris

2006

Biotechnol. Prog.

View full text Add to dashboard Cite

show abstract

“…At this point, cells entered the stationary phase. Other groups reported similar profiles for the specific antibody secretion rate in batch culture (34, 36, 56). Osmotic stress altered the secretion rate profile, reversing the steady decline.…”

Section: Resultsmentioning

confidence: 53%

“…To calculate the antibody assembly rate using the previously described model, eqs 1−3 were combined as described by McKinney et al (56): where…”

Section: Resultsmentioning

confidence: 99%

Hyperosmotic Stress in Murine Hybridoma Cells: Effects on Antibody Transcription, Translation, Posttranslational Processing, and the Cell Cycle

Sun

Zhou

Liang

et al. 2004

Biotechnology Progress

View full text Add to dashboard Cite

Mechanisms for increased antibody production in batch cultures of murine hybridoma cells in response to hyperosmotic stress were investigated. The rates of immunoglobulin transcription and protein translation and posttranslational processing were determined in control and hyperosmotic cultures. Changes in immunoglobulin transcription played a minor role in the increase in antibody production in response to hyperosmotic stress. In contrast, protein translation increased substantially in response to osmotic stress. However, the antibody translation rate remained relatively constant after correcting for the overall increase in protein translation. Cell size and intracellular antibody pool also increased in response to hyperosmolarity. The intracellular antibody pool increased proportionately with the increase in cell size, indicating that hyperosmotic cultures do not selectively increase their intracellular antibody population. Changes in cell cycle distribution in response to osmotic stress and the relationship between the cell cycle and antibody production were also evaluated. Hyperosmotic stress altered the cell cycle distribution, increasing the fraction of the cells in S-phase. However, this change was uncorrelated with the increase in antibody production rate. Immunoglobulin degradation was relatively low ( approximately 15%) and remained largely unchanged in response to hyperosmotic stress. There was no apparent increase in immunoglobulin stability as a result of osmotic stress. Antibody secretion rates increased approximately 50% in response to osmotic stress, with a commensurate increase in the antibody assembly rate. The rate of transit through the entire posttranslational processing apparatus increased, particularly for immunoglobulin light chains. The levels of endoplasmic reticulum chaperones did not increase as a fraction of the total cellular protein but were increased on a per cell basis as the result of an increase in total cellular protein. A difference in the interactions between the immunoglobulin heavy chains and BiP/GRP78 was observed in response to hyperosmotic conditions. This change in interaction may be correlated with the decrease in transit time through the posttranslational pathways. The increase in the posttranslational processing rate appears to be commensurate with the increase in antibody production in response to hyperosmotic stress.

show abstract

“…We have therefore focused on mAbs, which are essential tools for validation of proteins. 15–17) However, the commonly used hybridoma-based mAb production requires preparation of recombinant proteins as antigens and is laborious and time-consuming, 18–21) making it impractical for creating mAbs against many candidate proteins identified by proteomics-based analysis and forcing researchers to preferentially analyze proteins of their own interest. A phage antibody library system can rapidly produce mAbs against many antigens in vitro .…”

Section: Establishment Of Antibody Proteomics Technology: a High-thromentioning

confidence: 99%

Development of novel drug delivery systems using phage display technology for clinical application of protein drugs

Nagano

Tsutsumi

2016

Proceedings of the Japan Academy. Ser. B: Physical and Biologic

View full text Add to dashboard Cite

Attempts are being made to develop therapeutic proteins for cancer, hepatitis, and autoimmune conditions, but their clinical applications are limited, except in the cases of drugs based on erythropoietin, granulocyte colony–stimulating factor, interferon-alpha, and antibodies, owing to problems with fundamental technologies for protein drug discovery. It is difficult to identify proteins useful as therapeutic seeds or targets. Another problem in using bioactive proteins is pleiotropic actions through receptors, making it hard to elicit desired effects without side effects. Additionally, bioactive proteins have poor therapeutic effects owing to degradation by proteases and rapid excretion from the circulatory system. Therefore, it is essential to establish a series of novel drug delivery systems (DDS) to overcome these problems. Here, we review original technologies in DDS. First, we introduce antibody proteomics technology for effective selection of proteins useful as therapeutic seeds or targets and identification of various kinds of proteins, such as cancer-specific proteins, cancer metastasis–related proteins, and a cisplatin resistance–related protein. Especially Ephrin receptor A10 is expressed in breast tumor tissues but not in normal tissues and is a promising drug target potentially useful for breast cancer treatment. Moreover, we have developed a system for rapidly creating functional mutant proteins to optimize the seeds for therapeutic applications and used this system to generate various kinds of functional cytokine muteins. Among them, R1antTNF is a TNFR1-selective antagonistic mutant of TNF and is the first mutein converted from agonist to antagonist. We also review a novel polymer-conjugation system to improve the in vivo stability of bioactive proteins. Site-specific PEGylated R1antTNF is uniform at the molecular level, and its bioactivity is similar to that of unmodified R1antTNF. In the future, we hope that many innovative protein drugs will be developed by combining these technologies.

show abstract

Optimizing antibody production in batch hybridoma cell culture

Cited by 17 publications

References 59 publications

Development and Analysis of a Mathematical Model for Antibody-Producing GS-NS0 Cells Under Normal and Hyperosmotic Culture Conditions

Development and Analysis of a Mathematical Model for Antibody-Producing GS-NS0 Cells Under Normal and Hyperosmotic Culture Conditions

Hyperosmotic Stress in Murine Hybridoma Cells: Effects on Antibody Transcription, Translation, Posttranslational Processing, and the Cell Cycle

Development of novel drug delivery systems using phage display technology for clinical application of protein drugs

Contact Info

Product

Resources

About