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Integrated scale down of the primary downstream purification process and assessment of alternative process options for the production of an ovine polyclonal antibody based anti-venom.

Neal, G.E.; (2005) Integrated scale down of the primary downstream purification process and assessment of alternative process options for the production of an ovine polyclonal antibody based anti-venom. Doctoral thesis , University of London. Green open access

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Abstract

This thesis describes the development of a linked 1000th scale mimic of four distinct unit operations, including precipitation and centrifugation. These operations are utilised during the production of an anti-snake venom Fab fragment. A new approach has been used to define the important engineering parameters that impacted on the recovery process the effect of these was verified by experimental work. The mimic requires only millilitre quantities of material to predict changes in the physical and biological properties of key components, such as particle size and antibody activity. The potential impact of several process changes on the characteristics of the feed stream and on purification steps further downstream has been assessed, which is not possible without a linked system. Microfiltration has been examined as a possible alternative to the current centrifugal method of recovering the antibody particles. A small-scale stirred cell device was used to carry out a number of operations in a single piece of equipment these included separation, concentration and buffer exchange. An overall increase in the yield of 10% was observed. This was attributed to the ability of microfiltration to reduce material losses by integrating a number of operations. A preliminary investigation has been conducted into the possibility of utilising Protein G affinity chromatography in place of the current four-step precipitation and centrifugation recovery operation. A mathematical model capable of predicting the size and shape of the breakthrough curves has been developed the predicted curves were tested by comparison with breakthrough curves produced under different experimental conditions. The initial results demonstrated the feasibility of utilising affinity chromatography as a one step recovery process. However the predicted breakthrough curves varied from the experimental curves suggesting that the mathematical model requires refinement.

Type: Thesis (Doctoral)
Title: Integrated scale down of the primary downstream purification process and assessment of alternative process options for the production of an ovine polyclonal antibody based anti-venom.
Identifier: PQ ETD:592174
Open access status: An open access version is available from UCL Discovery
Language: English
Additional information: Thesis digitised by ProQuest
UCL classification: UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering
URI: https://discovery.ucl.ac.uk/id/eprint/1444864
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