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Recombinant protein expression by mammalian cells is often limited by the intracellular rate of post-translational carboxylation of the product protein. It is known that increasing the level of carbonic anhydrase (CA) activity and thus the intracellular concentration of free bicarbonate and thus CO2 in the culture media increases the production of glycosylated recombinant human growth hormone. A similar increase in glycosylation rate has been shown with mouse-derived recombinant human granulocyte colony stimulating factor. The present study was designed to determine whether these results could be replicated using the Chinese hamster ovary cell line, CHO-K1. The results show that recombinant human granulocyte colony stimulating factor and recombinant human growth hormone could be synthesised in CHO-K1 cells, but the protein was produced as a mixture of glycosylated and non-glycosylated forms. Upon increasing the concentration of bicarbonate in the culture media from 0 to 0.75% (v/v) the glycosylation of both recombinant growth factor and recombinant factor decreased from approximately 33 to 13% and approximately 23 to 14% respectively. Increasing the bicarbonate concentration of the growth media to 2.0% or higher only marginally enhanced the levels of glycosylated recombinant factor. Increasing the concentration of exogenous CO2 in the culture media from 10 to 25% increased the level of glycosylation of the recombinant growth factor by only 10%, but the level of glycosylation of the recombinant factor increased by approximately 23% for both acidic and basic isoforms. The results of this study indicate that the levels of recombinant factor that can be secreted in CHO cells can be increased by increasing the concentration of bicarbonate in the media to approximately 2% (v/v) but further increases in bicarbonate concentration will lead to levels of recombinant factor that are below the level that would be obtained with 1% bic
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The brainchild of toy and novel enthusiast Marcus Piggott, Anything But Ordinary is a daily updated blog featuring news, views and exclusive interviews from the worlds of toys, games and popular culture.On August 7, D.C. police “made an arrest, recovered drugs and cash, and seized personal belongings.”
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According to a report from Bloomberg, a consortium of crypto-lenders including Bitfinex, Galaxy Digital, and BitMEX led the $70m purchase of a 27.5% stake in a $800m loan to Bitfinex from the three UK-based lenders, LendInvest, Quorum, and Genesis Capital.
The deal was brokered by a new crypto-based bank called Aergo, created by Block Financial and their affiliate Cohava Labs, which will be the first crypto bank in Asia.
There’s also an investment fund, Aergo Capital, and a $35m strategic investment that has been confirmed as financing for the bank.
“We are excited to begin this journey with the first crypto bank in Asia and a partnership with LendInvest, Quorum, and Genesis Capital,” said Aergo’s head of communications and business development, Beau Stebbings. “It is an exciting time for our company and we look forward to continuing to build on the positive momentum for crypto that is currently underway worldwide.”
The deal follows an announcement from the LendInvest in May that it planned to buy 70% of the loan.
Steebbings continued: “We are seeing an upward trajectory for the crypto markets — prices have been relatively stable from highs to lows and gains in recent months are real and sustained. We look forward to supporting Bitfinex in this continued growth and leveraging Aergo’s platform and network to help Bit
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