Goodbye E.coli : Alternativa endotoxinfria produktionssystem till Escherichia coli

Sammanfattning: Affibody is interested in replacing Escherichia coli as the production system for Affibody®-molecules in order to reduce the amount of resources needed for the purification step. The aim of this report is to present relevant endotoxin-free alternatives that would suit production. Affibody®-molecules are proteins that have similar specificity as antibodies used in biopharmaceuticals, but have the advantage of being smaller. Since E. coli is the current production organism, the product is contaminated with immunogenic endotoxins which cause a high demand of resources during the purification process, something Affibody wants to reduce. The results presented in this report were achieved by means of a systematic literature study, divided into one general phase and two specific search phases. In the general literature search phase we were solely concerned with finding endotoxin-free expression systems. In the specific search phases the expression systems found were narrowed down in two steps based on criteria such as potential of scale-up > 1000 L, approval from the European Medicines Agency and the Food and Drug Administration, cost and productivity. Finally, only three candidates remained that met all requirements. The information found pertaining to the expression systems was presented in a spreadsheet, as were the different sources used. The nominated endotoxin-free candidates suggested to replace E. coli are Pichia pastoris, Chinese Hamster Ovary (CHO) cells and the Baculovirus Expression Vector System (BEVS). The candidates were then weighed against each other in an analysis based on the given criteria. We concluded that all three candidates satisfy the criteria and are suitable replacements for Affibody to use instead of E. coli.