PMID- 27666939
OWN - NLM
STAT- MEDLINE
DCOM- 20171017
LR  - 20171215
IS  - 1097-0290 (Electronic)
IS  - 0006-3592 (Linking)
VI  - 114
IP  - 3
DP  - 2017 Mar
TI  - FX knockout CHO hosts can express desired ratios of fucosylated or afucosylated 
      antibodies with high titers and comparable product quality.
PG  - 632-644
LID - 10.1002/bit.26188 [doi]
AB  - During antibody dependent cell cytotoxicity (ADCC) the target cells are killed by 
      monocytes and natural killer cells. ADCC is enhanced when the antibody heavy 
      chain's core N-linked glycan lacks the fucose molecule(s). Several strategies 
      have been utilized to generate fully afucosylated antibodies. A commonly used and 
      efficient approach has been knocking out the FUT8 gene of the Chinese hamster 
      ovary (CHO) host cells, which results in expression of antibody molecules with 
      fully afucosylated glycans. However, a major drawback of the FUT8-KO host is the 
      requirement for undertaking two separate cell line development (CLD) efforts in 
      order to obtain both primarily fucosylated and fully afucosylated antibody 
      species for comparative studies in vitro and in vivo. Even more challenging is 
      obtaining primarily fucosylated and FUT8-KO clones with similar enough product 
      quality attributes to ensure that any observed ADCC advantage(s) can be strictly 
      attributed to afucosylation. Here, we report generation and use of a FX knockout 
      (FXKO) CHO host cell line that is capable of expressing antibody molecules with 
      either primarily fucosylated or fully afucosylated glycan profiles with otherwise 
      similar product quality attributes, depending on addition of fucose to the cell 
      culture media. Hence, the FXKO host not only obviates the requirement for 
      undertaking two separate CLD efforts, but it also averts the need for screening 
      many colonies to identify clones with comparable product qualities. Finally, FXKO 
      clones can express antibodies with the desired ratio of primarily fucosylated to 
      afucosylated glycans when fucose is titrated into the production media, to allow 
      achieving intended levels of FcgammaRIII-binding and ADCC for an antibody. 
      Biotechnol. Bioeng. 2017;114: 632-644. (c) 2016 Wiley Periodicals, Inc.
CI  - (c) 2016 Wiley Periodicals, Inc.
FAU - Louie, Salina
AU  - Louie S
AD  - Early Stage Cell Culture, Genentech, Inc, 1 DNA Way, South San Francisco, 
      California, 94080.
FAU - Haley, Benjamin
AU  - Haley B
AD  - Department of Molecular Biology, Genentech, Inc, South San Francisco, California.
FAU - Marshall, Brett
AU  - Marshall B
AD  - Department of Analytical Development and Quality Control, Genentech, Inc, South 
      San Francisco, California.
FAU - Heidersbach, Amy
AU  - Heidersbach A
AD  - Department of Molecular Biology, Genentech, Inc, South San Francisco, California.
FAU - Yim, Mandy
AU  - Yim M
AD  - Early Stage Cell Culture, Genentech, Inc, 1 DNA Way, South San Francisco, 
      California, 94080.
FAU - Brozynski, Martina
AU  - Brozynski M
AD  - Department of Analytical Development and Quality Control, Genentech, Inc, South 
      San Francisco, California.
FAU - Tang, Danming
AU  - Tang D
AD  - Early Stage Cell Culture, Genentech, Inc, 1 DNA Way, South San Francisco, 
      California, 94080.
FAU - Lam, Cynthia
AU  - Lam C
AD  - Early Stage Cell Culture, Genentech, Inc, 1 DNA Way, South San Francisco, 
      California, 94080.
FAU - Petryniak, Bronislawa
AU  - Petryniak B
AD  - Department of Pathology, Genentech, Inc, South San Francisco, California.
FAU - Shaw, David
AU  - Shaw D
AD  - Early Stage Cell Culture, Genentech, Inc, 1 DNA Way, South San Francisco, 
      California, 94080.
FAU - Shim, Jeongsup
AU  - Shim J
AD  - Department of Analytical Development and Quality Control, Genentech, Inc, South 
      San Francisco, California.
FAU - Miller, Aaron
AU  - Miller A
AD  - Department of Analytical Development and Quality Control, Genentech, Inc, South 
      San Francisco, California.
FAU - Lowe, John B
AU  - Lowe JB
AD  - Department of Pathology, Genentech, Inc, South San Francisco, California.
FAU - Snedecor, Brad
AU  - Snedecor B
AD  - Early Stage Cell Culture, Genentech, Inc, 1 DNA Way, South San Francisco, 
      California, 94080.
FAU - Misaghi, Shahram
AU  - Misaghi S
AD  - Early Stage Cell Culture, Genentech, Inc, 1 DNA Way, South San Francisco, 
      California, 94080.
LA  - eng
PT  - Journal Article
DEP - 20161004
PL  - United States
TA  - Biotechnol Bioeng
JT  - Biotechnology and bioengineering
JID - 7502021
RN  - 0 (Antibodies)
RN  - 0 (Recombinant Proteins)
RN  - 28RYY2IV3F (Fucose)
RN  - EC 1.2.- (Ketone Oxidoreductases)
RN  - EC 1.2.1.- (GDP-4-keto-6-deoxy-D-mannose reductase)
SB  - IM
MH  - Animals
MH  - Antibodies/*chemistry/genetics/metabolism
MH  - CHO Cells
MH  - CRISPR-Cas Systems
MH  - Cricetinae
MH  - Cricetulus
MH  - Fucose/chemistry/*metabolism
MH  - Gene Editing
MH  - Gene Knockout Techniques
MH  - Ketone Oxidoreductases/*genetics
MH  - Protein Engineering/*methods
MH  - Recombinant Proteins/*chemistry/genetics/metabolism
OTO - NOTNLM
OT  - GDP-4-keto-6-d-deoxymannose epimerase/GDP-4-keto-6-l-galactose reductase (FX)
OT  - fucose
OT  - fucosylation
OT  - fully afucosylated antibody
OT  - glycans
EDAT- 2016/09/27 06:00
MHDA- 2017/10/19 06:00
CRDT- 2016/09/27 06:00
PHST- 2016/05/20 00:00 [received]
PHST- 2016/09/09 00:00 [revised]
PHST- 2016/09/23 00:00 [accepted]
PHST- 2016/09/27 06:00 [pubmed]
PHST- 2017/10/19 06:00 [medline]
PHST- 2016/09/27 06:00 [entrez]
AID - 10.1002/bit.26188 [doi]
PST - ppublish
SO  - Biotechnol Bioeng. 2017 Mar;114(3):632-644. doi: 10.1002/bit.26188. Epub 2016 Oct 
      4.