Rational surface design and process monitoring in biotechnology
Abstract
This thesis describes three approaches to create rational chromatographic surfaces. These surfaces exhibit specificities which are akin to that of antibodies. Thus they provide an alternative to using antibody columns for protein purification. Such surfaces do not suffer from the problems commonly associated with antibody columns like denaturation etc. as the purification is done under milder conditions. Yet they exhibit comparable specificity. The first three chapters describe in detail three separate approaches to generate such surfaces. Process monitoring is a very important issue in the biotechnology industry. While chapter 4 discusses the need for process monitoring and the technology available today to perform process monitoring, chapter 5 discusses different examples like monitoring of hybridomas, fermentors etc. It also describes assays which have been developed to separate and quantitate IgG from its aggregates. Use of digital filtering and ensemble averaging to increase signal to noise ratios, for process monitoring is also demonstrated. Chapter 6 and 7 discuss the fabrication of a new support which has C-8 chains on a polystyrene divinylbenzene backbone. This support exhibits unique selectivities and results in improved separation of small molecules, tryptic peptides and biomolecules. Finally in chapter 8 a newly developed immunological assay is described. This is an ELISA based assay which is performed in capillaries instead of traditional polystyrene wells. This reduces the volume and results in increased amplification of the enzyme, thus increasing sensitivity and decreasing assay time. Using this assay, it was possible to detect 300 zmol of human IgG in 20 min.
Degree
Ph.D.
Advisors
Regnier, Purdue University.
Subject Area
Analytical chemistry|Biochemistry|Molecular biology
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