Degludec

We have developed technology for secretory expression of human insulin precursor (IP) with alpha factor leader under methanol-inducible promoter from yeast Pichia pastoris. IP is purified and cleaved by trypsin in order to remove the N-terminal extension and the spacer tripeptide therefore generating the product DesB30. Finally, a hexadecanedioic acid is attached via gamma-L-glutamyl spacer to the amino group of the side chain of lysine at position B29 in order to obtain the long-lasting insulin Degludec.

 

Introduction video: Degludec

Video Procedures

1

Preparation of desB30-insulin

2

Preparation of DEGLUDEC synthetic linker composed of hexadecanedioic acid and a gamma-L-glutamyl spacer

3

Modification of DesB30-insulin with activated deprotected Degludec linker

4

Chromatographic purification of Degludec from the reaction mixture

To access the course please contact:

Natasa Skoko, Head

Biotechnology Development Unit

+39 040 3757352

skoko@icgeb.org

Martina Viviani, Head

Technology Transfer Office

+39 040 3757365

viviani@icgeb.org