* CHOgro
¢ç Expression Media, Dry Powder, Prepares 10 LitersÀÇ ÃÖ¼Ò ¹ßÁÖ¼ö·®Àº 3°³ ÀÌ»óÀ̸ç, ³³±â±îÁö 8~10ÁÖ ¼Ò¿äµË´Ï´Ù.
** CHOgro
¢ç Expression Media, 10 Liter PolybagÀÇ ÃÖ¼Ò ÁÖ¹® ¼ö·®Àº 10°³ ÀÌ»óÀ̸ç, ³³±â±îÁö 8~10ÁÖ ¼Ò¿äµË´Ï´Ù.
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CHOgro¢ç Expression SystemÀº suspension CHO CellÀ» ÀÌ¿ëÇÏ¿© ´ë·®ÀÇ ´Ü¹éÁú ¹ßÇö¿¡ ÃÖÀûÀÎ Á¦Ç°ÀÌ´Ù. Transfection ½Ã¾à°ú ¹èÁö°¡ ¸ðµÎ Æ÷ÇÔµÈ All-in-one kitÀ̸ç, ´Ù¸¥ culture systemÀ» »ç¿ëÇÏ°í ÀÖ´Â ¿¬±¸ÀÚµµ CHOgro Expression Medium ¿¡ 24½Ã°£ ÀûÀÀ ÈÄ TransfectionÀ» ÁøÇàÇϸé, °£ÆíÇÏ°Ô ³ôÀº ´Ü¹éÁú ¹ßÇöÀ» °æÇèÇÒ ¼ö ÀÖ´Ù.
- ¼Â¾÷ÀÌ °£Æí: ´ë·® ´Ü¹éÁú »ý»ê¿¡ ÃÖÀûÀÎ ÇÁ·ÎÅäÄÝ Á¦°ø
- Cell clumpingÀ» ÃÖ¼ÒÈÇÏ¿©, ³ôÀº ¹ÐµµÀÇ CHO cell ¹è¾ç (12x10^6 cells/ml) ¹× ¶Ù¾î³ viability ½ÇÇö
- Chemically defined, hydrolysate-free, animal origin free medium
- cGMP (±¹Á¦ÀǾàÇ° Ç°Áú ±âÁØ) Á¶°Ç¿¡¼ »ý»ê - CHOgro¢ç Expression Medium
CHOgro¢ç Expression System ±¸¼º
±¸¼ºÇ° |
¿ë·® |
º°µµ±¸¸Å °¡´É |
TransIT-PRO¢ç Transfection Reagent |
1 ml |
MIR 5740 |
CHOgro¢ç Expression Medium |
2 ¡¿ 1000 ml |
MIR 6200 |
CHOgro¢ç Complex Formation Solution |
100 ml |
MIR 6210 |
Poloxamer 188 Solution |
100 ml |
MIR 6230 |
L-Glutamine Solution |
100 ml |
MIR 6240 |
Media Á¾·ù¿¡ µû¸¥ ¼¼Æ÷ ¹è¾ç ¹Ðµµ ¹× viability ºñ±³
Figure 1. Suspension CHO cells grow to high density in the CHOgro¢ç Expression Medium. Tripli-cate flasks of FreeStyle¢â CHO-S cells were seeded in CHOgro¢ç
Expression Medium (red line) or FreeStyle¢â CHO Expression Medium (blue line) at a cell density of 0.5 x 106cells/§¢. Cell counts (solid line) and viability (dotted line) were measured
daily using a Guava easyCyte¢â 5HT flow cytometer.
Expression system¿¡ µû¸¥ ´Ü¹éÁú ¹ßÇö ºñ±³
Figure 2. CHOgro¢ç Expression Medium yields multi-fold increases in antibody titer. Human IgG1 was produced by transient transfection using TransIT-PRO¢ç (1:1),
FreeStyle¢â MAX (1.25:1.25) or 25 kDa linear PEI (6:1) transfection reagents according to the manufacturers¡¯ or published protocol (reagent:DNA ratio). FreeStyle¢â
CHO-S cells were transfected using 1§¶ plasmid DNA per milliliter of culture and cell densities of 2 x 106 cells/§¢ or 1 x 106 cells/§¢ for the CHOgro¢ç Expression Medium (red bars)
or FreeStyle¢â Expression Medium (blue bars). Antibody levels were also analyzed from day 6 clarified supernatants using a human IgG ELISA.
°£ÆíÇÑ culture system º¯°æ°ú ¹ßÇöÀ² Áõ°¡
Figure 3. Media exchange leads to higher protein production. Free-Style¢â CHO-S cells were cultured in FreeStyle¢â CHO Expression Medium or CHOgro¢ç Expression Medium and
24 hours prior to transfection a subset of the cells grown in FreeStyle¢â CHO Expression Medium were spun down and exchanged with 100% fresh CHOgro¢ç Expression Medium.
The cells were allowed to grow and adapt for 24 hours prior to transfection with FreeStyle¢â MAX or TransIT-PRO¢ç transfection reagents according to the manufacturers¡¯
protocol and a hIgG1 encoding construct. (A) Workflow schematic of media exchange of CHO-S cells from FreeStyle¢â CHO Expression Medium to CHOgro¢ç Expression Medium
(black arrow) or the normal CHOgro¢ç Expression System (red arrow) (B) Day 6 supernatants were clarified and analyzed using a human IgG ELISA. Data is normalized to the
complete CHOgro¢ç Expression System (red bar).
´ë¿ë·® (10L) CHOgro¢ç Expression Media
±×¸² 4. ´ë·®ÀÇ CHO cell ¹è¾çÀ» À§ÇÑ ´ë¿ë·®ÀÇ CHOgro¢ç Expression Media. Powder ÇüÅ (ÁÂ), Polybag ÇüÅ (¿ì)
Human IgG1 Expression Control
Æ÷À¯·ù ¼¼Æ÷¿¡¼ Ç×ü ¹ßÇö È®Àοë control
Figure 5. Human IgG1 Expression Control is an Optimized Premix of Heavy and Light Chain Constructs. The Human IgG1 Expression Control is an
equimolar premix of proprietary human IgG1 heavy and light chain expression constructs with antibody expression driven by CMV promoters. This product incorporates DNA2.0 IP-Free¨Ï vector technology.
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°í¹ßÇö transfection reagent TransIT-PRO¢ç Transfection Reagent Á¤º¸ ÀÚ¼¼È÷ º¸±â
Endotoxin Á¦°Å¸¦ ÅëÇÑ ´Ü¹éÁú ¹ßÇö Çâ»ó MiraCLEAN¢ç Endotoxin Removal Kit ÀÚ¼¼È÷º¸±â
Transfection È¿À² È®Àοë Label IT¢ç Plasmid Delivery Control, Cy¢ç 3 ÀÚ¼¼È÷º¸±â