PMID- 24939591
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20150822
LR  - 20211021
IS  - 0920-9069 (Print)
IS  - 1573-0778 (Electronic)
IS  - 0920-9069 (Linking)
VI  - 67
IP  - 5
DP  - 2015 Oct
TI  - Production of anti TNF-alpha antibodies in eukaryotic cells using different 
      combinations of vectors carrying heavy and light chains.
PG  - 761-72
LID - 10.1007/s10616-014-9714-3 [doi]
AB  - Tumor necrosis factor-alpha (TNF-alpha) plays a key role in rheumatoid arthritis and some 
      other autoimmune diseases. Therapy with anti-TNF-alpha recombinant antibodies (Ab) 
      appears to be highly effective. Production of new hyper-producing eukaryotic cell 
      lines can decrease the treatment cost, which currently is very high. However, due 
      to the complexity of protein transcription, translation, processing, and 
      secretion in mammalian cells, the stages at which antibody expression is affected 
      are still poorly determined. The aim of this work was to compare the productivity 
      of two cell lines developed in CHO DG44 cells, deficient in dihydrofolate 
      reductase, transfected with vectors carrying either heavy (H) or light (L) chains 
      of chimeric antibody under different combinations of selective elements. Both H 
      and L chains were cloned either in pOptiVEC or pcDNA3.3 vectors and different 
      combinations were used to produce HL and LH cell lines. We have shown that Ab 
      production has been low and comparable between HL and LH cells until selection on 
      methotrexate (MTX) when LH but not HL cells have responded with 3.5 times 
      increased productivity. Flow cytometry analysis has demonstrated that 
      intracellular concentration of full size Abs in LH cells was 5.6 times higher 
      than in HL ones due to higher amount of H chain synthesis. No differences in 
      viability between HL and LH cells have been found. We have concluded that the 
      expression of H chain in the pOptiVEC vector, which is responsible for MTX 
      resistance, has led to the suppression of H chain synthesis and limitation in 
      full Ab assembly.
FAU - Balabashin, Dmitriy
AU  - Balabashin D
AD  - Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, RAS, GSP-7, 
      Miklukho-Maklaya, 16/10, 117997, Moscow, Russia, dbalabashin@gmail.com.
FAU - Kovalenko, Elena
AU  - Kovalenko E
FAU - Toporova, Viktoria
AU  - Toporova V
FAU - Aliev, Teimur
AU  - Aliev T
FAU - Panina, Anna
AU  - Panina A
FAU - Svirshchevskaya, Elena
AU  - Svirshchevskaya E
FAU - Dolgikh, Dmitry
AU  - Dolgikh D
FAU - Kirpichnikov, Mikhail
AU  - Kirpichnikov M
LA  - eng
PT  - Journal Article
DEP - 20140618
PL  - United States
TA  - Cytotechnology
JT  - Cytotechnology
JID - 8807027
PMC - PMC4545435
EDAT- 2014/06/19 06:00
MHDA- 2014/06/19 06:01
PMCR- 2016/10/01
CRDT- 2014/06/19 06:00
PHST- 2013/08/23 00:00 [received]
PHST- 2014/03/03 00:00 [accepted]
PHST- 2014/06/19 06:00 [entrez]
PHST- 2014/06/19 06:00 [pubmed]
PHST- 2014/06/19 06:01 [medline]
PHST- 2016/10/01 00:00 [pmc-release]
AID - 9714 [pii]
AID - 10.1007/s10616-014-9714-3 [doi]
PST - ppublish
SO  - Cytotechnology. 2015 Oct;67(5):761-72. doi: 10.1007/s10616-014-9714-3. Epub 2014 
      Jun 18.