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A central composite design to investigate antibody fragment production by Pichia pastoris

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dc.contributor.author Chan, Joyce, M. Eng. Massachusetts Institute of Technology en_US
dc.contributor.other Massachusetts Institute of Technology. Biological Engineering Division. en_US
dc.date.accessioned 2006-08-25T18:52:51Z
dc.date.available 2006-08-25T18:52:51Z
dc.date.copyright 2005 en_US
dc.date.issued 2005 en_US
dc.identifier.uri http://hdl.handle.net/1721.1/33872
dc.description Thesis (M. Eng.)--Massachusetts Institute of Technology, Biological Engineering Division, 2005. en_US
dc.description Includes bibliographical references (p. 77-80). en_US
dc.description.abstract This study aims to investigate the relationships between growth parameters (agitation, glycerol concentration, salt concentration) and responses (biomass, growth rate, protein expression), by a 3-factor-3-level central composite factorial design. This experimental design involved running shake flask culture at 15 different experimental conditions with duplicates. Optical density (OD600), dry cell weight (DCW), and BCA Protein Assays were done on each experiment. Mathematical models in terms of these parameters' effects and their interactions were proposed for each of the responses. The significance of each effect and interaction, as well as the goodness-of-fit of mathematical models to data were examined by analysis of variance. It was found that biomass (with R²Adj=0.951) is a strong function of glycerol concentration (higher glycerol concentration leads to higher biomass), but it varies much less with agitation, and it is completely independent of salt concentration. Growth rate (R²Adj=0.901), however, varies strongly with agitation and salt concentration, but much more weakly with glycerol concentration. Protein production has a low R²Adj value of 0.746, implying that higher-order terms, e.g. x₁² and x₂², should be tested for significance in the model. en_US
dc.description.abstract (cont.) Collected data were fitted to the proposed models by response surface regression, after which surface and contour plots of responses were generated to identify trends in them. High agitation (300 rpm in shaker) gave rise to both highest biomass and growth rate. In addition, biomass at high glycerol concentration (3% v/v) was almost twice as much as biomass at low glycerol concentration (1% v/v) at high agitation rate (19 g/L compared to 11 g/L). At the same agitation rate, growth rate shows the largest increase of 20.5% with increasing salt concentration from 0.7% to 2.1%. Protein production reached maximum of 7.3 mg/mL at medium agitation rate (250 rpm), high salt and glycerol concentrations. en_US
dc.description.statementofresponsibility by Joyce Chan. en_US
dc.format.extent 80 p. en_US
dc.format.extent 3207400 bytes
dc.format.extent 3210658 bytes
dc.format.mimetype application/pdf
dc.format.mimetype application/pdf
dc.language.iso eng en_US
dc.publisher Massachusetts Institute of Technology en_US
dc.rights M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission. en_US
dc.rights.uri http://dspace.mit.edu/handle/1721.1/7582
dc.subject Biological Engineering Division. en_US
dc.title A central composite design to investigate antibody fragment production by Pichia pastoris en_US
dc.type Thesis en_US
dc.description.degree M.Eng. en_US
dc.contributor.department Massachusetts Institute of Technology. Biological Engineering Division. en_US
dc.identifier.oclc 66464394 en_US


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