High-level expression of recombinant IgG in the human cell line per.c6.
The number of therapeutic monoclonal antibodies in production is expected to rise rapidly in the next few years. As a result, there is much focus on the optimization of antibody expression platforms. Several issues are important including the speed of transition from bench to manufacturing, yield of IgG, and quality (particularly of the glycan structures present on immunoglobulins). We have characterized the human cell line PER.C6 for its ability to produce recombinant IgG. Production yields are still being optimized, but in nonfed batch culture, PER.C6 is able to grow to a cell density of 5 x 10(6) cells/mL and produce 300-500 mg/L IgG; this is likely to increase significantly in fed batch cultures. The generation of antibody-producing cell lines is fast, as rounds of amplification of inserted genes are not required for high production yields. The gene copy number of inserted genes is in the region of 1-10 copies per genome. In addition, PER.C6 is a human cell line, and so does not add glycans, which are immunogenic in humans. A core fucose molecule is essentially always present, and galactose residues are present at a physiological level (0, 1, and 2 galactose residues per glycan are present at a ratio of 1:2:1). No hybrid or high-mannose structures are seen.
Jones
David
D
Crucell, Archimedesweg 4, P.O. Box 2048, 2301 CA Leiden, The Netherlands.
Kroos
Nathalie
N
Anema
Regina
R
van Montfort
Bart
B
Vooys
Andre
A
van der Kraats
Sven
S
van der Helm
Esmeralda
E
Smits
Shirley
S
Schouten
Jan
J
Brouwer
Kirsten
K
Lagerwerf
Fija
F
van Berkel
Patrick
P
Opstelten
Dirk-Jan
DJ
Logtenberg
Ton
T
Bout
Abraham
A
D019149:Bioreactors; D002455:Cell Division / Q000235:genetics / Q000502:physiology; D002460:Cell Line; D002470:Cell Survival / Q000235:genetics / Q000502:physiology; D003001:Cloning, Molecular / Q000379:methods; D005786:Gene Expression Regulation / Q000235:genetics*; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis* / Q000235:genetics; D011134:Polysaccharides / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D011786:Quality Control; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D012160:Retina / Q000166:cytology / Q000196:embryology / Q000378:metabolism* / Q000502:physiology; D014162:Transfection / Q000379:methods
D007074:Immunoglobulin G; D011134:Polysaccharides; D011994:Recombinant Proteins