Hesketh JE

Khazaipoul S

Optun A

Pearson MJ

Pryme IF

Stern B

Chinese hamster ovary (CHO) cells are used for recombinant protein production in the pharmaceutical industry but there is a need to improve expression levels. In the present work experiments were carried out to test the effectiveness of different 3\u27untranslated regions (3\u27UTRs) in promoting production of a naturally secreted luciferase. Seamless cloning was used to produce expression vectors in which Gaussia princeps luciferase coding sequences were linked to the human albumin, immunoglobulin or chymotrypsinogen 3\u27UTR. Stably transfected CHO cells expressing these constructs were selected. Luciferase activity in the culture medium was increased 23-fold by replacing the endogenous 3\u27UTR with the albumin 3\u27UTR and decreased by replacement with immunoglobulin or chymotrypsinogen 3\u27UTR. Replacement of the native 3\u27UTR with the albumin 3\u27UTR led to a 10-fold increase in luciferase mRNA levels. Deletion analysis of the albumin 3\u27UTR showed that loss of nucleotides 1-50, which removed an AU-rich complex stem loop region, caused significant reductions in both luciferase protein expression and luciferase mRNA levels. The results suggest that recombinant protein expression and yield could be improved by the careful selection of appropriate 3\u27UTR sequences

Albumin 3\u27untranslated region facilitates increased recombinant protein production from Chinese hamster ovary cells

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