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MAbs. 2013 Jul-Aug;5(4):595-607. doi: 10.4161/mabs.25161. Epub 2013 May 31.

Expression of antibodies using single open reading frame (sORF) vector design: Demonstration of manufacturing feasibility.

Author information

1
AbbVie, Worcester, MA, USA. wendy.gion@abbvie.com

Abstract

Efficient production of large quantities of therapeutic antibodies is becoming a major goal of the pharmaceutical industry. We developed a proprietary expression system using a polyprotein precursor-based approach to antibody expression in mammalian cells. In this approach, the coding regions for heavy and light chains are included within a single open reading frame (sORF) separated by an in-frame intein gene. A single mRNA and subsequent polypeptide are produced upon transient and stable transfection into HEK293 and CHO cells, respectively. Heavy and light chains are separated by the autocatalytic action of the intein and antibody processing proceeds to produce active, secreted antibody. Here, we report advances in sORF technology toward establishment of a viable manufacturing platform for therapeutic antibodies in CHO cells. Increasing expression levels and improving antibody processing by intein and signal peptide selection are discussed.

KEYWORDS:

antibody expression; intein; manufacturing; signal peptide; single open reading frame

PMID:
23774760
PMCID:
PMC3906313
DOI:
10.4161/mabs.25161
[Indexed for MEDLINE]
Free PMC Article
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