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Home > ÀüÁ¦Ç°º¸±â > Gene transfer (virus) > Lentivirus > RetroNectin¢ç Pro (Recombinant Human Fibronectin Fragment)
G-Rex¢ç Bioreactor applicationÀÌ Ãß°¡µÈ, T-cell expansion ¹× transduction enhancer

RetroNectin¢ç Pro (Recombinant Human Fibronectin Fragment)

-

Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
Takara
T101A
RetroNectin¢ç Pro (Recombinant Human Fibronectin Fragment)
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
0.5 §· (0.5 §¢)
240,000¿ø  T101A_T101B_DS.v2409.pdf
Takara
T101B
RetroNectin¢ç Pro (Recombinant Human Fibronectin Fragment)
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
2.5 §· (2.5 §¢)
894,000¿ø 

  • °¡½º Åõ°ú¼º ¿ë±â (G-Rex¢ç Bioreactor ½Ã¸®Áî) application Ãß°¡, ºü¸£°í Æí¸®ÇÑ transduction ¹× T-cell expansion °¡´É
  • ±âÁ¸ RetroNectin¢ç (Code T100A)°ú ¼º´ÉÀº ±×´ë·Î (µ¿ÀÏÇÑ ÇÁ·ÎÅäÄÝ) °¡°ÝÀº ´Ù¿î
  • Lentivirus¿Í RetrovirusÀÇ Transduction ÃËÁø
  • Naive T-cell ºñÀ²ÀÌ ³ôÀº È®´ë¹è¾ç ÇÁ·ÎÅäÄÝ
Á¦Ç°¼³¸í
RetroNectin¢ç Pro (Recombinant Human Fibronectin Fragment)´Â human fibronectinÀÇ ÀçÁ¶ÇÕ ´Ü¹éÁú*·Î ¼¼°¡ÁöÀÇ domain (cell binding domain; C-domain / heparin binding domain; H-domain, CS-1 domain)À» Æ÷ÇÔÇÏ°í ÀÖ´Ù (±×¸² 1). Lentivirus ¹× RetrovirusÀÇ transduction½Ã »ç¿ëÇϸé VLA-4 ¶Ç´Â VLA-5¸¦ ¹ßÇöÇÏ´Â ¼¼Æ÷°¡ °¢°¢ CS-1 domain°ú C-domain (RGDS ¼­¿­)¿¡ ºÎÂøµÇ¸ç, ¹ÙÀÌ·¯½º°¡ H-domain¿¡ °áÇյȴÙ. À̶§ ¹ÙÀÌ·¯½º¿Í ¸ñÀû ¼¼Æ÷ÀÇ ¹°¸®Àû °Å¸®°¡ ÃÖ¼ÒÈ­µÇ´Â ¿ø¸®·Î transductionÀÌ ÃËÁøµÈ´Ù.
¶ÇÇÑ T-cellÀÇ È®´ë¹è¾ç½Ã interleukin 2 (IL-2)ÀÇ Á¸Àç ÇÏ¿¡¼­ Anti-CD3 mAb GMP grade (Code T210)¿Í ÇÔ²² »ç¿ëÇϸé T-cellÀ» ÀÚ±ØÇØ naive T-cellÀ» °íÈ¿À²·Î È®´ë¹è¾çÇÒ ¼ö ÀÖ´Ù.

*ºÐÀÚ·®: 62,617 Da (¾Æ¹Ì³ë»ê ¼­¿­ ±âÁØ)

±×¸² 1. Fibronectin°ú RetroNectin¢çÀÇ ±¸Á¶ ¹× À¯ÀüÀÚ Àü´Þ ¸ðµ¨ ´ÙÀ̾î±×·¥
°¡½ºÅõ°ú¼º ¹è¾ç ¿ë±â (G-Rex 24 well plate)¸¦ »ç¿ëÇÏ¿© T-cell·Î À¯ÀüÀÚ Àü´Þ ¹× È®´ë¹è¾ç
°¡½º Åõ°ú¼º ¹è¾ç ¿ë±â (G-Rex¢ç Bioreactor ½Ã¸®Áî)´Â °í¹Ðµµ ¹è¾çÀÌ °¡´ÉÇÏ¿© ¼¼Æ÷ÀÇ È®´ë¹è¾ç¿¡ ÀÚÁÖ »ç¿ëµÇ´Â ½Ã½ºÅÛÀÌ´Ù. À̶§ lentivirus µîÀ¸·Î T-cell¿¡ À¯ÀüÀÚ¸¦ Àü´ÞÇÏ´Â °æ¿ì, ¾Æ·¡ ±×¸²2¿Í °°ÀÌ plate µî¿¡¼­ T-cellÀÇ activation ºÎÅÍ À¯ÀüÀÚ Àü´Þ ±îÁöÀÇ °úÁ¤À» ÁøÇàÇÏ°í ¼¼Æ÷¸¦ G-Rex·Î ¿Å°Ü¼­ È®´ë¹è¾çÇÑ´Ù (Âü°í¹®Çå ÂüÁ¶). º» ¿¬±¸¿¡¼­´Â À§ÀÇ ¹æ¹ý ´ë½Å RetroNectin¢ç Pro¿Í anti-CD3 Ç×ü¸¦ G-Rex vessel¿¡ Á÷Á¢ ÄÚÆÃÇÏ°í, ÄÚÆÃÇÑ G-Rex vessel¿¡¼­ À¯ÀüÀÚ Àü´Þ ¹× Áõ½Ä ¹è¾çÀ» ÁøÇàÇÏ¿© naive T cellÀÇ À¯ÀüÀÚ Àü´Þ È¿À² ¹× Áõ½Ä È¿°ú¸¦ °ËÁõÇß´Ù.

Âü°í¹®Çå:
Mapping variant effects on anti-tumor hallmarks of primary human T cells with base-editing screens
Zachary H.Walsh, Parin Shah, Neeharika Kothapalli, et al., Nature Biotechnology (2024)

±×¸² 2. G-Rex 24 well plateÀÇ À¯ÀüÀÚ Àü´Þ ¹× È®´ë¹è¾ç ¹æ¹ý
RetroNectin¢çÀ» »ç¿ëÇÏÁö ¾ÊÀº °æ¿ìÀÇ ºñ±³ Å×½ºÆ®
- ½ÇÇè Á¶°Ç

½ÇÇè Á¶°Ç

A

B

PlateÀÇ ÄÚÆÃ

RetroNectin¢ç Pro¿Í Anti-CD3 mAb GMP grade (#T210)¸¦ 4¡É¿¡¼­ O/Nµ¿¾È ÄÚÆÃÇÑ ÈÄ ÀÜ¿©¹° Á¦°Å ¹× ¼¼Ã´

-

T-cell activation

CD3/CD28 bead »ç¿ë

¼¼Æ÷

CD8 positive ¹× CD4 ¾ç¼º T ¼¼Æ÷

cell ¼ö

1¡¿106 cells

¹è¾ç ºÎÇÇ

1.0 §¢

Transduction ½ÃÀÛ ½ÃÁ¡

¹è¾ç ½ÃÀÛÀÏ (Day 0)

Lentivirus

CD19-CAR À¯ÀüÀÚ¸¦ Àü´ÞÇÏ´Â Lentivirus¸¦ MOI 5¿Í 15·Î °¨¿°

¹è¾ç ¹èÁö

LymphoONE¢ç T-Cell Expansion Xeno-Free Medium, 1L Bottle (Code WK552S)¿¡ human serum albumin°ú IL-2¸¦ ÷°¡


- °á°ú

±×¸² 3. T-cell expansion ÈÄ cell ¼ö
10ÀÏ µ¿¾È ¹è¾ç ÈÄ ¼¼Æ÷ 1 well (2 cm)2 ´ç cell ¼öÀÌ´Ù. G-Rex ½Ã½ºÅÛÀ» »ç¿ëÇϸé A¿Í B ¹æ¹ý ¸ðµÎ °í¹Ðµµ·Î ¹è¾ç (2x107 cell/cm2 ÀÌ»ó)ÇÒ ¼ö ÀÖÀ½À» È®ÀÎÇß´Ù.


±×¸² 4. À¯ÀüÀÚ Àü´Þ È¿À²
Transduction ÈÄ 4ÀÏÂ÷¿Í 10ÀÏÂ÷¿¡ Å×½ºÆ®ÇÑ ¼¼Æ÷ÀÇ CAR ¹ßÇöÀ²À» ºÐ¼®Çß´Ù. ±× °á°ú, RetroNection¢ç Pro Á¶°Ç (A)¿¡¼­ ´õ¿í ³ôÀº ¹ßÇöÀ²À» º¸¿´°í CAR ¹ßÇöÀ²ÀÌ ³ôÀº ¼¼Æ÷°¡ ¾ò¾îÁö´Â °ÍÀ» È®ÀÎÇß´Ù.

±×¸² 5. Transduction ÈÄ naive T-cell ºñÀ²
TransductionÈÄ 10ÀÏÂ÷¿¡ naive T-cellÀÇ ºñÀ²À» ºÐ¼®Çß´Ù. ±× °á°ú, CD8+CAR+ ¹× CD4+CAR+ ¼¼Æ÷ÀÇ naive T-cell ºñÀ²ÀÌ MOI (15 ¶Ç´Â 5)¿Í °ü°è¾øÀÌ RetroNectin¢ç Pro ó¸®Á¶°Ç (A)¿¡¼­ ´õ ³ôÀº °ÍÀ» È®ÀÎÇß´Ù.

À§ÀÇ ¼¼°¡ÁöÀÇ µ¥ÀÌÅ͸¦ ÅëÇØ G-Rex¿¡ RetroNectin¢ç Pro ¿Í anti-CD3 Ç×ü¸¦ ÄÚÆÃÇÔÀ¸·Î½á ¹ÙÀÌ·¯½º¿¡ ÀÇÇÑ À¯ÀüÀÚ Àü´Þ°ú ÀÌÈÄÀÇ T-cellÀÇ È®´ë¹è¾çÀ» µ¿½Ã¿¡ ÁøÇàÇÒ ¼ö ÀÖ°í, ³ôÀº transduction È¿À²°ú ´Ù·®ÀÇ naive T-cellÀ» È®º¸ÇÒ ¼ö ÀÖÀ½À» ¾Ë ¼ö ÀÖ´Ù.

G-Rex´Â ÇÊ¿ä½Ã °£´ÜÇÏ°Ô scale-upÀÌ °¡´ÉÇÑ ½Ã½ºÅÛÀ̹ǷΠRetroNectin¢çÀ» °°ÀÌ »ç¿ëÇϸé ÀÛ¾÷ È¿À²À» Å©°Ô Çâ»ó½ÃÄÑ ¡°cost effective¡±ÇÑ ½ÇÇèÀ» ÁøÇàÇÒ ¼ö ÀÖÀ» °ÍÀ¸·Î ±â´ëÇÑ´Ù.
Çü»ó
ÀÌ Á¦Ç°Àº ¿©°ú ¹× ¸ê±ÕµÈ »óÅÂÀÇ solution (1 §·/§¢) ÀÌ´Ù.
(Á¦Çü) 12.5 mM sodium citrate (pH 6.2), 1.25% sucrose
º¸Á¸ -20¡É
±â¿ø
Human fibronectin fragment CH-296¸¦ ¹ßÇöÇÏ´Â E. coli
³óµµ 1 §·/§¢
ºÐÀÚ·® 62,617 Da (amino acid sequence)
¼øµµ >90% by HPLC
»ç¿ë»óÀÇ ÁÖÀÇ
- ¸ê±ÕµÈ ȯ°æ¿¡¼­ »ç¿ëÇÑ´Ù.
- Freeze/Thaw´Â ÃÖ´ë 10ȸ±îÁö °¡´ÉÇÏ´Ù.
- °úµµÇÑ ±³¹Ý ¹× vortexingÀ» ÇÇÇÑ´Ù.
- RetroNectin¢ç Pro fragment°¡ ¼Õ½ÇµÉ ¼ö Àֱ⠶§¹®¿¡ PBS¿¡ Èñ¼®µÈ RetroNectin¢ç Pro solutionÀ» filter·Î ¸ê±ÕÇÏÁö ¸¶½Ã¿À.
°ü·ÃÁ¦Ç°
- T-cell È®´ë¹è¾ç ¿ë GMP grade antibody Anti-CD3 mAb GMP grade (Code T210)
- Xeno-Free, Serum-FreeÀÇ T-cell ¹è¾ç¹èÁö LymphoONE¢ç T-Cell Expansion Xeno-Free Medium, 1L Bottle (Code WK552S)
- ³ôÀº °¨¿° È°¼ºÀ» °¡Áö´Â AAV/lentivirus »ý»êÀ» À§ÇÑ transfection ½Ã¾à TransIT-VirusGEN¢ç Transfection Reagent (Code MIR 6700) (GMP gradeµµ ÆǸŠÁß)
- Mycoplasma ¿À¿° °ËÃâ, luminometer¸¦ »ç¿ëÇÑ MycoAlert¢â PLUS Mycoplasma detection kit (Code LT07-701)
- Mycoplasma ¿À¿° °ËÃâ, nested PCRÀ» »ç¿ëÇÑ TaKaRa PCR Mycoplasma Detection Set (Code 6601)
- Mycoplasma ¿À¿° °ËÃâ, Real-Time PCRÀ» »ç¿ëÇÑ CycleavePCR¢ç Mycoplasma Detection Kit (Code CY232)




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