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Förslaget inkom 2003-11-14

The effect of viral clearance techniques on protein quality

OBS! ANSÖKNINGSTIDEN FÖR DETTA EXJOBB HAR LÖPT UT.
Background
Regulatory authorities demand viral clearance evaluation of all recombinant protein processes cultivated from mammalian cells. The purification process shall include viral clearance steps proven to reduce or inactivate certain levels of virus. In most cases several viral clearance steps, based on different mechanisms, are incorporated into the purification process. Techniques to adequately inactivate or reduce viruses are often harsh and the effect on the target protein can in many cases be negative. A fast and uncomplicated method, which sufficiently inactivates or reduces viruses without effecting the target protein at all, is desired.

Aim
This work aims to evaluate known virus inactivating and reducing techniques and study their effect on three model proteins. Focus will be on the protein and the effect on protein quality, i.e. function, structure, stability and aggregation. The goal is to bring knowledge and hands-on experience of various viral clearance techniques and which protein quality effects these different techniques may have. The results from this thesis work will hopefully lay the ground for a study set up to use in development and optimisation of viral clearance techniques in future projects.

Practical work and Techniques
The practical work involves setting up the viral clearance methods and running them on the three different proteins. Different parameters will be studied in order to achieve an optimised set up for each protein. The effect on the protein quality will be monitored.

Viral clearance methods:
 Chemical virus inactivation, such as low pH and solvent detergent methods
 Virus filtration, various filters and suppliers
 Additional virus inactivation techniques of interest
Protein quality methods:
 Biophysical methods for analysis of stability and aggregation
 Protein structural analysis methods such as SDS-PAGE, IEF and Peptide map

Applicants
For this development project we are looking for a student with an education within the biotech area with a large interest of protein chemistry. The assignment will include experiment planning, laboratory work in purification and characterisation labs with the newest equipment, evaluation and interpretation of obtained data. The assignment is based in the Purification and Characterisation department, Biopharmaceutical development unit at Biovitrum in Stockholm.


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