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Studies to prevent degradation of recombinant fc-fusion protein expressed in mammalian cell line and protein characterization

Chakrabarti, Sanjukta, Barrow, Colin J., Kanwar, Rupinder K., Ramana, Venkata and Kanwar, Jagat R. 2016, Studies to prevent degradation of recombinant fc-fusion protein expressed in mammalian cell line and protein characterization, International journal of molecular sciences, vol. 17, no. 6, pp. 1-22, doi: 10.3390/ijms17060913.

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Title Studies to prevent degradation of recombinant fc-fusion protein expressed in mammalian cell line and protein characterization
Author(s) Chakrabarti, Sanjukta
Barrow, Colin J.ORCID iD for Barrow, Colin J. orcid.org/0000-0002-2153-7267
Kanwar, Rupinder K.
Ramana, Venkata
Kanwar, Jagat R.ORCID iD for Kanwar, Jagat R. orcid.org/0000-0003-3728-9568
Journal name International journal of molecular sciences
Volume number 17
Issue number 6
Article ID 913
Start page 1
End page 22
Total pages 22
Publisher MDPI
Place of publication Basel, Switzerland.
Publication date 2016-06
ISSN 1422-0067
Keyword(s) CHOK1SV GS-KO
Fc fusion protein
VEGFR1(D1–D3)-Fc
biological activity
clipping
folded
inhibition
proteolysis
stable
Science & Technology
Life Sciences & Biomedicine
Physical Sciences
Biochemistry & Molecular Biology
Chemistry, Multidisciplinary
Chemistry
VEGFR1(D1-D3)-Fc
CIRCULAR-DICHROISM SPECTROSCOPY
HAMSTER OVARY CELLS
HETEROGENEITY
PRODUCTIVITY
ANTIBODIES
PEPTIDES
Summary Clipping of recombinant proteins is a major issue in animal cell cultures. A recombinant Fc-fusion protein, VEGFR1(D1-D3)-Fc expressed in CHOK1SV GS-KO cells was observed to be undergoing clippings in lab scale cultures. Partial cleaving of expressed protein initiated early on in cell culture and was observed to increase over time in culture and also on storage. In this study, a few parameters were explored in a bid to inhibit clipping in the fusion protein The effects of culture temperature, duration of culture, the addition of an anti-clumping agent, ferric citrate and use of protease inhibitor cocktail on inhibition of proteolysis of the Fc fusion were studied. Lowering of culture temperature from 37 to 30 °C alone appears to be the best solution for reducing protein degradation from the quality, cost and regulatory points of view. The obtained Fc protein was characterized and found to be in its stable folded state, exhibiting a high affinity for its ligand and also biological and functional activities.
Language eng
DOI 10.3390/ijms17060913
Field of Research 119999 Medical and Health Sciences not elsewhere classified
0399 Other Chemical Sciences
0604 Genetics
0699 Other Biological Sciences
Socio Economic Objective 929999 Health not elsewhere classified
HERDC Research category C1 Refereed article in a scholarly journal
ERA Research output type C Journal article
Copyright notice ©2016, The Authors
Free to Read? Yes
Use Rights Creative Commons Attribution licence
Persistent URL http://hdl.handle.net/10536/DRO/DU:30085982

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Every reasonable effort has been made to ensure that permission has been obtained for items included in DRO. If you believe that your rights have been infringed by this repository, please contact drosupport@deakin.edu.au.