title: Expression of antibodies using single open reading frame (sORF) vector design: Demonstration of manufacturing feasibility.
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.
authors.affiliation: AbbVie, Worcester, MA, USA. wendy.gion@abbvie.com
authors.forename: Wendy R
authors.initials: WR
authors.lastname: Gion
authors.forename: Rachel A
authors.initials: RA
authors.lastname: Davis-Taber
authors.forename: Dean A
authors.initials: DA
authors.lastname: Regier
authors.forename: Emma
authors.initials: E
authors.lastname: Fung
authors.forename: Limary
authors.initials: L
authors.lastname: Medina
authors.forename: Ling C
authors.initials: LC
authors.lastname: Santora
authors.forename: Sahana
authors.initials: S
authors.lastname: Bose
authors.forename: Alexander V
authors.initials: AV
authors.lastname: Ivanov
authors.forename: Barbara A
authors.initials: BA
authors.lastname: Perilli-Palmer
authors.forename: Chris M
authors.initials: CM
authors.lastname: Chumsae
authors.forename: Joseph G
authors.initials: JG
authors.lastname: Matuck
authors.forename: Yune Z
authors.initials: YZ
authors.lastname: Kunes
authors.forename: Gerald R
authors.initials: GR
authors.lastname: Carson
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000096:biosynthesis / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000096:biosynthesis / Q000235:genetics; D047668:Inteins; D016366:Open Reading Frames; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D057127:Single-Chain Antibodies / Q000096:biosynthesis / Q000235:genetics
chemical_list: D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies
keywords: antibody expression; intein; manufacturing; signal peptide; single open reading frame