Cloning, expression, cellcultivation and purification of a mutant of the enzyme human arylsulfatase A (hASA)

To whom: This project is suitable for students within Biotechnology, Biomedicine, Cell and Molecular Biology, Biochemistry etc. The student should have knowledge and experience in general laboratory work within the field and 2-4 years of relevant university studies

The aim of this project is to perform cloning and expression work of a specific mutant of rhASA (the cDNA and also CHO cells are available as starting points of this project). When a pool of CHO cells expressing the rhASA mutant is available, cell propagation in shakers will be the next step. The rhASA mutant will be secreted in the medium and purification of the enzyme will take place. This can be achieved by either conventional chromatography (3 steps) or antibody based affinity chromatography.

This mutant will then be used as a research tool to link specific peptides that that will be tested for their eventual capacity to facilitate transfer of the enzyme over the blood brain barrier in order to improve the effect on the CNS (in collaboration with univeristy groups in Europe)

This project will give a good insight and also hands on experience of almost all steps involved in laboratory scale biotechnology projects today. The project has an inbuilt flexibility since depending on success rate the project can be either shortened or expanded.