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Kinetics study of hybridoma growth and antibody production Henry, Olivier
Abstract
Monoclonal Antibodies (MAb) produced by hybridomas have an expanding market for use in diagnostic assays, affinity separations and therapeutic applications. The objectives of this thesis were to study the kinetics of growth and antibody production of a hybridoma cell line in five different modes of operation and establish to what extent these kinetics are comparable from one mode to another. Hybridoma cells producing monoclonal antibodies were grown in batch, fed-batch, semi-continuous, continuous and perfusion cultures. The kinetics were compared in terms of growth rate, nutrient consumption rate and antibody formation rate. Batch cultures reached a maximum cell concentration of around 3x106 cells/mL and antibody concentration of 100 |j,g/mL. Glutamine was likely the substrate limiting the batch cultures since its depletion coincided with the end of exponential growth phase. No other amino acid was consumed completely and the levels of toxic metabolites (23 mM lactate; 2.6 mM ammonium) were much lower than those reported to affect growth or MAb production. A simple unstructured model was developed based on the batch data during the exponential phase. Simulations were performed and compared with experimental results obtained in semicontinuous and continuous cultures at dilution rates varying from 0.016 to 0.036 h"1. Cell yields from glucose in batch exponential phase were approximately 30% less than in the continuous processes. The cell specific glucose and glutamine uptake rates increased at greater dilution rates in both continuous and daily fed semi-continuous cultures. Comparing the cultures on the basis of specific dilution rates, perfusion results were in good agreement with semi-continuous and continuous data in terms of glutamine and glucose uptake rates. Although the specific antibody production rate only varied within a narrow range (0.7 to 2 pg/cell.h), the data suggest that the formation of antibody was growth-associated. It is concluded that modeling and analysis of semi-continuous data can describe continuous processes sufficiently well to be a useful tool for perfusion system optimization, while providing the relative simplicity of batch operation. In fed-batch, through the daily addition of a concentrate feed, cultures yielded antibody concentrations two times greater (200 (Xg/mL) compared to batch. This increase in final antibody concentration was closely related to the increase in the viable cell index.
Item Metadata
Title |
Kinetics study of hybridoma growth and antibody production
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Creator | |
Publisher |
University of British Columbia
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Date Issued |
2000
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Description |
Monoclonal Antibodies (MAb) produced by hybridomas have an expanding market for use in
diagnostic assays, affinity separations and therapeutic applications. The objectives of this
thesis were to study the kinetics of growth and antibody production of a hybridoma cell line
in five different modes of operation and establish to what extent these kinetics are
comparable from one mode to another. Hybridoma cells producing monoclonal antibodies
were grown in batch, fed-batch, semi-continuous, continuous and perfusion cultures. The
kinetics were compared in terms of growth rate, nutrient consumption rate and antibody
formation rate.
Batch cultures reached a maximum cell concentration of around 3x106 cells/mL and antibody
concentration of 100 |j,g/mL. Glutamine was likely the substrate limiting the batch cultures
since its depletion coincided with the end of exponential growth phase. No other amino acid
was consumed completely and the levels of toxic metabolites (23 mM lactate; 2.6 mM
ammonium) were much lower than those reported to affect growth or MAb production. A
simple unstructured model was developed based on the batch data during the exponential
phase. Simulations were performed and compared with experimental results obtained in semicontinuous
and continuous cultures at dilution rates varying from 0.016 to 0.036 h"1. Cell
yields from glucose in batch exponential phase were approximately 30% less than in the
continuous processes. The cell specific glucose and glutamine uptake rates increased at
greater dilution rates in both continuous and daily fed semi-continuous cultures. Comparing
the cultures on the basis of specific dilution rates, perfusion results were in good agreement with semi-continuous and continuous data in terms of glutamine and glucose uptake rates.
Although the specific antibody production rate only varied within a narrow range (0.7 to 2
pg/cell.h), the data suggest that the formation of antibody was growth-associated. It is
concluded that modeling and analysis of semi-continuous data can describe continuous
processes sufficiently well to be a useful tool for perfusion system optimization, while
providing the relative simplicity of batch operation.
In fed-batch, through the daily addition of a concentrate feed, cultures yielded antibody
concentrations two times greater (200 (Xg/mL) compared to batch. This increase in final
antibody concentration was closely related to the increase in the viable cell index.
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Extent |
4220551 bytes
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Genre | |
Type | |
File Format |
application/pdf
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Language |
eng
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Date Available |
2009-07-06
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Provider |
Vancouver : University of British Columbia Library
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Rights |
For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.
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DOI |
10.14288/1.0058764
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URI | |
Degree | |
Program | |
Affiliation | |
Degree Grantor |
University of British Columbia
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Graduation Date |
1999-11
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Campus | |
Scholarly Level |
Graduate
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Aggregated Source Repository |
DSpace
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Item Citations and Data
Rights
For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.