Abstract：
: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.
journal_name
Int J Mol Scijournal_title
International journal of molecular sciencesauthors
Chakrabarti S,Barrow CJ,Kanwar RK,Ramana V,Kanwar JRdoi
10.3390/ijms17060913subject
Has Abstractpub_date
2016-06-09 00:00:00issue
6issn
1422-0067pii
ijms17060913journal_volume
17pub_type
杂志文章abstract：:Melanocytes (MCs) are specialized cells that synthesize melanin within the melanosome. Cultured MCs are useful in order to study their role in relation to pigmentation. However, MC isolation is laborious and the obtained cells have a limited culture time. In this study, we transformed lentivirus-mediated simian virus ...
journal_title：International journal of molecular sciences
pub_type： 杂志文章
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abstract：:Neonatal seizures are one of the most common comorbidities of neonatal encephalopathy, with seizures aggravating acute injury and clinical outcomes. Current treatment can control early life seizures; however, a high level of pharmacoresistance remains among infants, with increasing evidence suggesting current anti-sei...
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journal_title：International journal of molecular sciences
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journal_title：International journal of molecular sciences
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更新日期：2012-01-01 00:00:00
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journal_title：International journal of molecular sciences
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更新日期：2019-09-09 00:00:00
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更新日期：2018-12-10 00:00:00
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更新日期：2019-09-10 00:00:00
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journal_title：International journal of molecular sciences
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更新日期：2018-10-30 00:00:00
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abstract：:The protein phosphatase PP2A is essential for the control of integrated eukaryotic cell functioning. Several cellular and developmental events, e.g., plant growth regulator (PGR) mediated signaling pathways are regulated by reversible phosphorylation of vesicle traffic proteins. Reviewing present knowledge on the rele...
journal_title：International journal of molecular sciences
pub_type： 杂志文章,评审
doi：10.3390/ijms22020975
更新日期：2021-01-19 00:00:00
