Development of mammalian production cell lines expressing CNTO736, a glucagon like peptide-1-MIMETIBODY: factors that influence productivity and product quality.
In an attempt to develop a high producing mammalian cell line expressing CNTO736, a Glucagon like peptide-1-antibody fusion protein (also known as a Glucagon like peptide-1 MIMETIBODY), we have noted that the N-terminal GLP-1 portion of the MIMETIBODY was susceptible to proteolytic degradation during cell culture, which resulted in an inactive product. Therefore, a number of parameters that had an effect on productivity as well as product quality were examined. Results suggest that the choice of the host cell line had a significant effect on the overall product quality. Product expressed in mouse myeloma host cell lines had a lesser degree of proteolytic degradation and variability in O-linked glycosylation as compared to that expressed in CHO host cell lines. The choice of a specific CHOK1SV derived clone also had an effect on the product quality. In general, molecules that exhibited minimal N-terminal clipping had increased level of O-linked glycosylation in the linker region, giving credence to the hypothesis that O-linked glycosylation acts to protect against proteolytic degradation. Moreover, products with reduced potential for N-terminal clipping had longer in vivo serum half-life. These findings suggest that early monitoring of product quality should be an essential part of production cell line development and therefore, has been incorporated in our process of cell line development for this class of molecules.
Dorai
Haimanti
H
Centocor R&D, 145 King of Prussia Road, Radnor, Pennsylvania 19087, USA. hdorai@its.jnj.com
Nemeth
Jennifer F
JF
Cammaart
Erwin
E
Wang
Yonghui
Y
Tang
Qing Mike
QM
Magill
Allen
A
Lewis
Michael J
MJ
Raju
T Shantha
TS
Picha
Kristen
K
O'Neil
Karyn
K
Ganguly
Subinay
S
Moore
Gordon
G
D000818:Animals; D001709:Biotechnology / Q000379:methods*; D002460:Cell Line; D006224:Cricetinae; D006801:Humans; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*
C534787:CNTO 736; D011993:Recombinant Fusion Proteins