Packages
Work Package 9
Enzyme production of human LAMAN
- Work Package Number: 9
- Involves: Partner 2a
The human recombinant LAMAN enzyme has been cloned and expressed by Zymenex. In order to provide sufficient amounts of the enzyme for the animal studies it is necessary to develop the process and produce 0.5 grams of enzyme. The enzyme is produced in CHO mammalian cells and the development will encompass establishing and increasing the cell fermentation capacity from shaker flasks level to 5 liter perfusion fermentors. The fermented cell extract is then purified using purification methods that will be developed and will require increasing the purification columns sizes from 20 ml to 200 to 1000 ml scale. The purified bulk must then be analyzed using assays that must be developed: 1) an activity assay for LAMAN activity, 2) SDS-PAGE, 3) reverse-phase HPLC and 4) BCA-protein determination. The final product must be formulated and filled in such a way, that it can be dosed to the mice.