albumin - publications

Predict more albumin - ligand interactions now!


1. Biotechnol J. 2012 May 23. doi: 10.1002/biot.201200044. [Epub ahead of print]

Albumin 3'untranslated region facilitates increased recombinant protein
production from Chinese Hamster Ovary Cells.

Pearson MJ, Khazaipoul S, Optun A, Pryme IF, Stern B, Hesketh JE.

Institute for Cell and Molecular Biosciences, The Medical School, Newcastle
University, Framlington Place, Newcastle upon Tyne, NE2 4HH, UK.

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'untranslated regions (UTRs) in promoting production of a naturally
secreted luciferase. Seamless cloning was used to produce expression vectors in
which G. princeps luciferase coding sequences were linked to the human albumin,
immunoglobulin or chymotrypsinogen 3'UTR. Stably transfected CHO cells expressing
these constructs were selected. Luciferase activity in the culture medium was
increased 2-3 fold by replacing the endogenous 3'UTR with the albumin 3'UTR and
decreased by replacement with immunoglobulin or chymotrypsinogen 3'UTR.
Replacement of the native 3'UTR with the albumin 3'UTR led to a 10-fold increase
in luciferase mRNA levels. Deletion analysis of the albumin 3'UTR 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'UTR
sequences.

Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

PMID: 22623341 [PubMed - as supplied by publisher]