Show simple item record

dc.contributor.advisorJ. Christopher Love.en_US
dc.contributor.authorMatthews, Catherine Bartlett.en_US
dc.contributor.otherMassachusetts Institute of Technology. Department of Chemical Engineering.en_US
dc.date.accessioned2019-10-11T22:00:25Z
dc.date.available2019-10-11T22:00:25Z
dc.date.copyright2019en_US
dc.date.issued2019en_US
dc.identifier.urihttps://hdl.handle.net/1721.1/122528
dc.descriptionThesis: Ph. D. in Chemical Engineering Practice, Massachusetts Institute of Technology, Department of Chemical Engineering, 2019en_US
dc.descriptionCataloged from PDF version of thesis.en_US
dc.descriptionIncludes bibliographical references.en_US
dc.description.abstractAdoption of non-mammalian host organisms for biologic drug manufacturing could lead to step-changes in cost of manufacturing and volumetric productivity, increasing access to these life-saving drugs for large patient populations. One promising alternative host is Pichia pastoris, a methylotrophic yeast that is currently used to manufacture ten approved drugs worldwide. Its fast growth rate and ability to grow to high cell densities can enable fast production cycles, agile process development, and potentially low production costs. While P. pastoris has already been engineered to produce antibodies with human-like glycoforms, titers are still lower than those typically achieved with CHO cells. While standard fermentation processes for P. pastoris have been designed, several areas have limited investment to date. Few chemically-defined cultivation media have been reported for P. pastoris fermentation and all are minimal salt solutions.en_US
dc.description.abstractWhile several studies have demonstrated that addition of complex nutrients improves growth and productivity, defined compounds with similar effects have not been identified. Also, methanol feeding protocols for P. pastoris have only been developed for fed-batch operation and have not been studied for perfusion cultivation. In this thesis, we describe the design of a rich defined medium (RDM) for cultivation of P. pastoris through systematic screening and gene expression analysis. The use of RDM for expression of three proteins yields titers comparable to, or higher than, those in standard complex medium. We then outline a similar methodology for the optimization of individual amino acids and fatty acids in the medium. We also describe how a transcriptomic analysis of methanol feeding strategy in perfusion mode enabled the identification and alleviation of limiting biological processes.en_US
dc.description.abstractThis work demonstrates how combining traditional process development strategies with genome-wide sequencing for P. pastoris leads to rapid improvement of fermentation processes. Continued progress in this area could lead to a new model for low-cost production of high-quality biologic drugs.en_US
dc.description.statementofresponsibilityby Catherine Bartlett Matthews.en_US
dc.format.extent136 pagesen_US
dc.language.isoengen_US
dc.publisherMassachusetts Institute of Technologyen_US
dc.rightsMIT theses are protected by copyright. They may be viewed, downloaded, or printed from this source but further reproduction or distribution in any format is prohibited without written permission.en_US
dc.rights.urihttp://dspace.mit.edu/handle/1721.1/7582en_US
dc.subjectChemical Engineering.en_US
dc.titleDesign of a cultivation medium for protein production in Pichia pastoris based on genome-wide biological understandingen_US
dc.typeThesisen_US
dc.description.degreePh. D. in Chemical Engineering Practiceen_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Chemical Engineeringen_US
dc.identifier.oclc1121594217en_US
dc.description.collectionPh.D.inChemicalEngineeringPractice Massachusetts Institute of Technology, Department of Chemical Engineeringen_US
dspace.imported2019-10-11T22:00:24Zen_US
mit.thesis.degreeDoctoralen_US
mit.thesis.departmentChemEngen_US


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record