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McMaster University

1. Sadavarte, Hemant Rahul. MEMBRANE AND TEMPERATURE BASED METHODS FOR PROCESSING AND PURIFYING MONOCLONAL ANTIBODIES.

Degree: MASc, 2013, McMaster University

Monoclonal antibodies (mAbs) as therapeutic proteins have shown great potential in treatment of various human diseases because of their highly specific nature. This has attracted worldwide attention leading to increased demand for such mAb products. To meet this demand large scale manufacturing is carried out using recombinant mammalian cell culture techniques for high yields and faster production. mAb products are worth the investment if produced in their native state. The quantity of mAb present in such cell cultures is very less and therefore special care is needed while handling them. Purifying antibody molecules from heterogeneous cell culture impurities and maintaining their native functional state is a critical task mainly because these antibodies are labile in nature. Care also need to be exercised during processing because mAbs have inherent tendancy to aggregate which is undesirable since such aggregates in antibody formulation produces immunogenic reaction when injected in humans. The other important factor in mAb purification is the processing cost involved since majority of the total production cost is utilized for purification of mAb. Protein-A chromatography is the first choice for purifying antibodies and is widely adopted. However failure in distinguishing between monomer and aggregate antibody molecules along with harsh acidic processing conditions necessitates the use of further purification steps. In this work various techniques for mAb processing are discussed and are outlined below: Removal of impurities from mAbs is a major challenge and this thesis discusses various processing options available to purify these mAbs. Impurities in mAb products are usually the aggregate byproducts formed due to unfolded monomer antibody molecules. These molecules are naturally hydrophobic in nature and display great differences in hydrophobicity on aggregation. Hydrophobic interaction membrane chromatography (HIMC) makes use of this hydrophobicity difference and helps in removal of aggregate impurities from monomer antibody. Heavy chain mAbs (hcmAbs) are promising new developments in the area of biopharmaceuticals because of their unique structural composition. Similar to conventional mAbs these hcmAbs are also rapidly finding their way into therapeutic markets. Purifying hcmAbs will be an important step in their development and for this purpose we use HIMC technique for removing impurities and obtain pure product. Antibody molecules are almost always lost as aggregates which leads to great economic losses and the ability to disaggregate these mAb oligomers would be of significant practical and scientific interest. In this work a novel thermalcycling technique is discussed to disaggregate such mAb oligomers and potentially recover functional monomer mAb molecules.

Master of Applied Science (MASc)

Advisors/Committee Members: Ghosh, Raja, Prashant Mhaskar, Ravi Selvaganapathy, Chemical Engineering.

Subjects/Keywords: Hydrophobic Interaction Membrane Chromatography; Thermal cycling of Monoclonal Antibodies; Chimeric Heavy Chain Monoclonal Antibody (cHCmAb); Processing and Purification of Monoclonal Antibodies; Membrane Adsorbers; IgG1; EG2.; Amino Acids, Peptides, and Proteins; Biotechnology; Macromolecular Substances; Membrane Science; Other Chemical Engineering; Amino Acids, Peptides, and Proteins

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Sadavarte, H. R. (2013). MEMBRANE AND TEMPERATURE BASED METHODS FOR PROCESSING AND PURIFYING MONOCLONAL ANTIBODIES. (Masters Thesis). McMaster University. Retrieved from http://hdl.handle.net/11375/12863

Chicago Manual of Style (16th Edition):

Sadavarte, Hemant Rahul. “MEMBRANE AND TEMPERATURE BASED METHODS FOR PROCESSING AND PURIFYING MONOCLONAL ANTIBODIES.” 2013. Masters Thesis, McMaster University. Accessed November 12, 2019. http://hdl.handle.net/11375/12863.

MLA Handbook (7th Edition):

Sadavarte, Hemant Rahul. “MEMBRANE AND TEMPERATURE BASED METHODS FOR PROCESSING AND PURIFYING MONOCLONAL ANTIBODIES.” 2013. Web. 12 Nov 2019.

Vancouver:

Sadavarte HR. MEMBRANE AND TEMPERATURE BASED METHODS FOR PROCESSING AND PURIFYING MONOCLONAL ANTIBODIES. [Internet] [Masters thesis]. McMaster University; 2013. [cited 2019 Nov 12]. Available from: http://hdl.handle.net/11375/12863.

Council of Science Editors:

Sadavarte HR. MEMBRANE AND TEMPERATURE BASED METHODS FOR PROCESSING AND PURIFYING MONOCLONAL ANTIBODIES. [Masters Thesis]. McMaster University; 2013. Available from: http://hdl.handle.net/11375/12863

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