High yield heterologous expression of human Intestinal Alkaline Phosphatase
Secretion of hiAP
After 48 h of fermentation, hiAP activities of 14,000 U/l were reached. The high yield could be confirmed by SDS gel after Coomassie stain and immunoblot.
The human Alkaline Phosphatase, which is under investigation for treatment against sepsis, was produced with very good yield. With a C-terminal truncated version without GPI-anchor, up to 14.000 U/l were reached, while other expression systems achieved considerably lower yields. A full length version with GPI-anchor was accumulated in the outer cortex of the Tetrahymena cell.
Localisation of full length hiAP with GPI-anchor
Detection of hiAp with fluorescent labeled antibodies on the surface of the cells.