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Application of the novel and convenient IR/MAR gene amplification technology to the production of recombinant protein pharmaceuticals.

Araki Y, Noguchi C, Hamafuji T, Nose H, Miki D, Shimizu N - BMC Proc (2011)

View Article: PubMed Central - HTML - PubMed

Affiliation: Graduate School of Biosphere Science, Hiroshima University, Higashi-hiroshima, 739-8521, Japan.

No MeSH data available.


The IR/MAR plasmid can generates DMs (A), HSR (B), Ladder-HSR (C), and Fine ladder-HSR (D). Among the metaphase chromosome spread, we detected the plasmid sequence by FISH in green. Gene expression was generally higher from DMs than HSR. However, DMs are usually hard to be generated in CHO cells. The IR/MAR plasmid can efficiently generate the ladder and the fine ladder structure that is active in transcription.
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Figure 1: The IR/MAR plasmid can generates DMs (A), HSR (B), Ladder-HSR (C), and Fine ladder-HSR (D). Among the metaphase chromosome spread, we detected the plasmid sequence by FISH in green. Gene expression was generally higher from DMs than HSR. However, DMs are usually hard to be generated in CHO cells. The IR/MAR plasmid can efficiently generate the ladder and the fine ladder structure that is active in transcription.

Mentions: As a result, the usage of IR/MAR technology enabled us to obtain cells, in which the introduced genes were amplified to a few hundreds to thousands copies per cells as DMs or HSR of various size and shape (Figure 1), which depended both on the vector constructs and the host cell lines. Such stable cells with amplified genes could be obtained within one month, and the protein production was increased more than a hundred-fold compared with the case without IR/MAR. A cell clone showed the specific production rate that reached almost the highest reported for antibody protein (45 pg/cell/day). Furthermore, we have found several novel ways that further improve the protein production level. For example, the combination of the IR/MAR and the DHFR/MTx technologies synergistically work and far more rapidly and easily generate the cells of higher production rate than previously.


Application of the novel and convenient IR/MAR gene amplification technology to the production of recombinant protein pharmaceuticals.

Araki Y, Noguchi C, Hamafuji T, Nose H, Miki D, Shimizu N - BMC Proc (2011)

The IR/MAR plasmid can generates DMs (A), HSR (B), Ladder-HSR (C), and Fine ladder-HSR (D). Among the metaphase chromosome spread, we detected the plasmid sequence by FISH in green. Gene expression was generally higher from DMs than HSR. However, DMs are usually hard to be generated in CHO cells. The IR/MAR plasmid can efficiently generate the ladder and the fine ladder structure that is active in transcription.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3285013&req=5

Figure 1: The IR/MAR plasmid can generates DMs (A), HSR (B), Ladder-HSR (C), and Fine ladder-HSR (D). Among the metaphase chromosome spread, we detected the plasmid sequence by FISH in green. Gene expression was generally higher from DMs than HSR. However, DMs are usually hard to be generated in CHO cells. The IR/MAR plasmid can efficiently generate the ladder and the fine ladder structure that is active in transcription.
Mentions: As a result, the usage of IR/MAR technology enabled us to obtain cells, in which the introduced genes were amplified to a few hundreds to thousands copies per cells as DMs or HSR of various size and shape (Figure 1), which depended both on the vector constructs and the host cell lines. Such stable cells with amplified genes could be obtained within one month, and the protein production was increased more than a hundred-fold compared with the case without IR/MAR. A cell clone showed the specific production rate that reached almost the highest reported for antibody protein (45 pg/cell/day). Furthermore, we have found several novel ways that further improve the protein production level. For example, the combination of the IR/MAR and the DHFR/MTx technologies synergistically work and far more rapidly and easily generate the cells of higher production rate than previously.

View Article: PubMed Central - HTML - PubMed

Affiliation: Graduate School of Biosphere Science, Hiroshima University, Higashi-hiroshima, 739-8521, Japan.

No MeSH data available.