
¡Ø RetroNectin¢ç GMP grade (Code T202) Á¦Ç° ÁÖ¹®½Ã
Àü¿ë ÁÖ¹®¼¸¦ ÀÛ¼ºÇÏ¿© ´ÙÄ«¶óÄÚ¸®¾Æ(
support@takara.co.kr)·Î º¸³»ÁÖ¼¼¿ä.
¡Ø µ¿°á°ÇÁ¶ ÇüÅÂÀÇ RetroNectin¢ç GMP grade (Code T201, Á¾¸Å)ÀÌ solution ÇüÅ (Code T202) ·Î ´ëüµÇ¾ú½À´Ï´Ù.
- Lentivirus ¿Í retrovirusÀÇ transduction enhancer
- Polybrene ¶Ç´Â protamineÀÇ Àû¿ëÀÌ ¾î·Á¿î ¼¼Æ÷µµ ³·Àº µ¶¼ºÀ¸·Î °íÈ¿À² transduction ½ÇÇö
- Hematopoietic cell°ú °°Àº suspension cellµµ °íÈ¿À² transduction °¡´É
- T-cell È®´ë¹è¾ç¿¡µµ Àû¿ëÇÒ ¼ö ÀÖ¾î CAR-T cell ¿¬±¸¿¡ ÃÖÀû
- ±âÁ¸ T-cell È®´ë¹è¾ç¹ý ´ëºñ ¾ÐµµÀûÀÎ È®´ë¹è¾ç
- T-cell È®´ë¹è¾ç Àû¿ë ¿¹: ±¹¹® (Korean)/ ¿µ¹® (English)
Á¦Ç°¼³¸í
RetroNectin® (Recombinant Human Fibronectin CH-296)Àº ¨ç Cell-binding domain (C-domain, VLA-5¿Í °áÇÕ) / ¨è Heparin-binding domain (H-domain, ¹ÙÀÌ·¯½º¿Í °áÇÕ) / ¨é CS-1 (VLA-4¿Í °áÇÕ), ÃÑ 3Á¾·ùÀÇ ±â´É¼º ´Ü¹éÁú ´ÜÆíÀ» °¡Áø human fibronectinÀÌ´Ù (±×¸² 1). ¸ñÀû ¼¼Æ÷°¡ VLA-4 ¶Ç´Â VLA-5¸¦ ¹ßÇöÇÏ¸é °¢°¢ CS-1 ¶Ç´Â Cell-binding domain°ú °áÇÕÇϰí À̶§ H-domain¿¡ °áÇÕÇÑ ¹ÙÀÌ·¯½º¿Í ¸ñÀû ¼¼Æ÷ÀÇ ¹°¸®Àû °Å¸®°¡ ÃÖ¼ÒȵǴ ¿ø¸®·Î transductionÀÌ ÃËÁøµÈ´Ù.

±×¸² 1. Fibronectin°ú RetroNectin®ÀÇ ±¸Á¶ ¹× transduction ¸ð½Äµµ
ÀϹÝÀûÀ¸·Î Hematopoietic cells ¶Ç´Â ±× ¿ÜÀÇ suspension ¼¼Æ÷µéÀº transduction È¿À²ÀÌ ³·Áö¸¸ RetroNectin®Àº ³·Àº µ¶¼ºÀ¸·Î ÀÌ·± ¼¼Æ÷¿¡¼µµ °íÈ¿À² transductionÀÌ °¡´ÉÇÏ´Ù. ÇöÀç, RetroNectin®Àº hematopoietic cellsÀÇ lentiviral / retroviral transduction¿¡ gold standard·Î »ç¿ëµÇ´Â enhancerÀ̸ç, 40°Ç ÀÌ»óÀÇ ÀÓ»ó ½ÃÇè¿¡ »ç¿ëµÇ¾î ÀÓ»ó Àû¿ë¿¡µµ Å« ±â´ë¸¦ ¹Þ°í ÀÖ´Ù.
¸ñÀû¼¼Æ÷·ÎÀÇ µµÀÔ¿¡´Â (1) supernatant¹ý ¶Ç´Â (2) RBV (RetroNectin-bound virus)¹ýÀ» Àû¿ëÇÒ ¼ö ÀÖ´Ù.
(1) Supernatant ¹ý
¼¼Æ÷¿Í ¹ÙÀÌ·¯½º¸¦ È¥ÇÕÇÏ¿© RetroNectin® ÄÚÆÃ Ç÷¹ÀÌÆ®¿¡ µµÀÔÇÏ´Â ¹æ¹ýÀ¸·Î ´Ü½Ã°£¿¡ °£ÆíÇÏ°Ô transductionÀ» ÁøÇàÇÒ ¼ö ÀÖ´Ù (¸ñÀû ¼¼Æ÷ ÷°¡ ÈÄ plate¸¦ ¿ø½ÉºÐ¸®ÇÏ¸é ¸ñÀû ¼¼Æ÷¿Í ¹ÙÀÌ·¯½ºÀÇ »óÈ£ÀÛ¿ëÀ» ÃËÁøÇÒ ¼ö ÀÖ´Ù.).

(2) RBV (RetroNectin-bound virus) ¹ý
¹ÙÀÌ·¯½º ¿ë¾×¿¡ packaging cell-line¿¡¼ ºÐºñµÈ proteoglycanÀ̳ª envelope proteinÀÌ È¥ÀԵǴ °æ¿ì transductionÀ» ÀúÇØÇÏ´Â °æ¿ì°¡ ÀÖ´Ù. RBV¹ýÀº ÀÌ·¯ÇÑ ÀúÇØ¹°ÁúÀ» Á¦°ÅÇϰí transductionÇÏ´Â ¹æ¹ýÀÌ´Ù. RetroNectin® ÄÚÆÃ Ç÷¹ÀÌÆ®¿¡ lentivirus ¶Ç´Â retrovirus¸¦ ¸ÕÀú °áÇÕ½Ã۰í transduction inhibitor°¡ È¥ÀÔµÈ ¹èÁö¸¦ Á¦°ÅÇÑ ÈÄ, ¸ñÀû¼¼Æ÷¸¦ ÷°¡ÇÏ¿© transductionÇÑ´Ù. Supernatant¹ýÀ¸·Î ÃæºÐÇÑ transduction È¿À²ÀÌ ³ª¿ÀÁö ¾ÊÀ» °æ¿ì RBV¹ýÀ» ÃßõÇÏ¸ç °í³óµµ·Î ¹ÙÀÌ·¯½º¸¦ »ç¿ëÇÏ´Â °æ¿ì¿¡¼ ³ôÀº È¿À²À» º¸Àδ٠(¹ÙÀÌ·¯½ºÀÇ titer°¡ ºÎÁ·ÇÑ °æ¿ì ¹ÙÀÌ·¯½º ¿ë¾× ÷°¡ ÈÄ plate¸¦ ¿ø½ÉºÐ¸®ÇÏ¸é ¹ÙÀÌ·¯½ºÀÇ transduction È¿À²À» ´õ¿í ³ôÀÏ ¼ö ÀÖ´Ù.).

* RetroNectin® Precoated Dish (Code T110A)´Â RetroNectin®ÀÌ 35 mm dish¿¡ ¹Ì¸® ÄÚÆÃµÈ ready-to-use dishÀÌ´Ù. ÄÚÆÃ°úÁ¤À» »ý·«Çϰí ÀÏÁ¤ÇÑ À¯ÀüÀÚ µµÀÔ È¿À²À» ¾òÀ» ¼ö ÀÖ´Ù.
* RetroNectin® GMP (Code T202)´Â GMP °¡À̵å¶óÀο¡ ÁØÇÑ Á¦Á¶ ¹× ǰÁú °ü¸® °øÁ¤¿¡ ÀÇÇØ »ý»êµÈ GMP gradeÀÇ RetroNectin®À¸·Î ¿¬±¸¿ëÀ¸·Î ÆÇ¸ÅµÇ´Â RetroNectin® (Code T100A/B)º¸´Ù °íǰÁú·Î À¯ÀüÀÚÄ¡·á³ª ¼¼Æ÷ Á¶Á¦ µîÀÇ ÀÓ»ó ¿¬±¸¿¡ »ç¿ë °¡´ÉÇÏ´Ù.
¿ëµµ
- LA-4 ¶Ç´Â VLA-5¸¦ ¹ßÇöÇÏ´Â ¼¼Æ÷ (´Ù¾çÇÑ hematopoietic cell Æ÷ÇÔ)¿¡ lentivirus ¶Ç´Â retrovirusÀÇ µµÀÔ(Transduction enhancer)
- T-cell (CD8+) È®´ë¹è¾ç
Àû¿ë ¿¹

±×¸² 2. ´Ù¾çÇÑ Áٱ⼼Æ÷¿¡ ´ëÇÑ retrovirusÀÇ À¯ÀüÀÚ µµÀÔ È¿À² ºñ±³.
´Ù¾çÇÑ Áٱ⼼Æ÷¿¡ ¼¼ °³ÀÇ transduction enhancer¸¦ ÀÌ¿ëÇÏ¿© retrovirus¸¦ »ç¿ëÇÑ Çü±¤ ´Ü¹éÁúÀÇ ¹ßÇö È¿À²À» ºñ±³ÇÏ¿´´Ù. ±× °á°ú, RetroNectin®Àº GALV envelope receptorÀÇ ¹ßÇöÀÌ ³·Àº hCD34+, hADSC °°Àº Áٱ⼼Æ÷¿¡¼µµ ³ôÀº È¿À²ÀÇ transductionÀÌ °¡´ÉÇÏ¿´´Ù.
Cell
|
Efficiency of Gene Transfer (%)
|
TF-1
|
53.3
|
HEL
|
28.1
|
NIH/3Y3
|
80.5
|
CD34+bMC
|
86.9
|
Mice BM MNC fraction
|
29.2
|
K-562
|
93.5
|
HL-60
|
89.4
|
Ç¥ 1. RetroNectin®À» »ç¿ëÇÑ transduction efficiency ¿¹½Ã

±×¸² 3. Lentivirus¸¦ »ç¿ëÇÑ suspension cell (SUP-T1) À¯ÀüÀÚ µµÀÔ »ç·Ê
suspension cellÀÎ SUP-T1¿¡ lentivirus¸¦ »ç¿ëÇÑ Çü±¤ ´Ü¹éÁúÀÇ ¹ßÇöÀ» ÀÌ¿ëÇÏ¿© transduction ¹æ¹ý º° È¿À²À» ºñ±³ÇÏ¿´´Ù. ±× °á°ú RetroNectin®À» »ç¿ëÇÑ Supernatant¹ý (static¹ý, spin¹ý), RBV-Spin¹ý ¸ðµÎ polybreneº¸´Ù ³ôÀº È¿À²·Î transductionÀÌ °¡´ÉÇÏ¿´´Ù.
Çü»ó
RetroNectin® (Recombinant Human Fibronectin Fragment) (Code T100A/B)
|
- 20¡É
|
RetroNectin® Dish (35 mm diameter Dish) (Code T110A)
|
4¡É
|
RetroNectin® GMP grade (Recombinant Human Fibronectin Fragment CH-296) (Code T202)
|
- 30¡É ÀÌÇÏ
|
±â¿ø
Human fibronectin fragment CH-296À» ¹ßÇöÇÏ´Â E.coli
¼øµµ
HPLC¿¡¼ ¼øµµ 90% ÀÌ»ó.
»ç¿ë »óÀÇ ÁÖÀÇ
- Freeze/thaw´Â 10ȸ* Á¤µµ±îÁö °¡´ÉÇÏ´Ù.
* RetroNectin® GMP grade (Code T202)´Â Çѹø ¿ëÇØ ÈÄ room temperature¿¡¼ 12½Ã°£ À̳» ¸ðµÎ »ç¿ëµÇ¾ß ÇÑ´Ù.
- °ú°ÝÇÑ ±³¹ÝÀº ÇÇÇÑ´Ù.
ºñ°í
RetroNectin®Àº lentivirus ¶Ç´Â retrovirusÀÇ À¯ÀüÀÚ µµÀÔ È¿À² Áõ°¡ »Ó¸¸ ¾Æ´Ï¶ó, T cellÀÇ È®´ë¹è¾ç¿¡µµ Àû¿ë °¡´ÉÇÔÀÌ È®ÀεǾú´Ù. ÀϹÝÀûÀ¸·Î T cell È®´ë ¹è¾çÀº Anti-CD3 antibody (OKT3)ÀÇ coating ÈÄ ¹è¾ç¾×¿¡ IL-2¸¦ ÷°¡ÇÏ¿© ½Ç½ÃÇÑ´Ù. À̶§, RetroNectin®À» Anti-CD3 antibody¿Í ÇÔ²² coatingÇÏ¸é ±âÁ¸ÀÇ È®´ë ¹è¾ç¹ý ´ëºñ ¾à 1.5¹è ÀÌ»ó È®´ë ¹è¾çÀÌ °¡´ÉÇÏ°í ³ôÀº naive T cell ºñÀ²À» À¯ÁöÇÒ ¼ö ÀÖ´Ù. ÀÌ·¯ÇÑ ¹æ¹ýÀº CAR-T cell ¿¬±¸¿¡ Gold standard·Î »ç¿ëµÇ°í ÀÖ´Ù.
°ü·ÃÁ¦Ç°
- ³ôÀº °¨¿° Ȱ¼ºÀ» °¡Áö´Â AAV/lentivirus »ý»êÀ» À§ÇÑ transfection ½Ã¾à TransIT-VirusGEN® Transfection Reagent (Code MIR6700) (GMP gradeµµ ÆÇ¸Å Áß)
- Mycoplasma ¿À¿° °ËÃâ, luminometer¸¦ »ç¿ëÇÑ MycoAlert 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® Mycoplasma Detection Kit (Code CY232)
Citation
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Generation of tumor-targeted human T lymphocytes from induced pluripotent stem cells for cancer therapy
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Stable expression of shRNAs in human CD34+ progenitor cells can avoid induction of interferon responses to siRNAs in vitro.
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ED SUM: The efficacy of chimeric antigen receptor (CAR)-T cells in solid tumor models is enhanced by IL23 engineering.
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A new cloning and expression system yields and validates TCRs from blood lymphocytes of patients with cancer within 10 days
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Heparanase promotes tumor infiltration and antitumor activity of CAR-redirected T-lymphocytes
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Correction of X-linked chronic granulomatous disease by gene therapy, augmented by insertional activation of MDS1-EVI1, PRDM16 or SETBP1.
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Safety and feasibility of anti-CD19 CAR T cells with fully-human binding domains in patients with B-cell lymphoma
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Immune recognition of somatic mutations leading to complete durable regression in metastatic breast cancer
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Immunogenicity and therapeutic targeting of a public neoantigen derived from mutated PIK3CA
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Anti-GD2 CAR-NKT cells in patients with relapsed or refractory neuroblastoma: an interim analysis
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Calibration of CAR activation potential directs alternative T cell fates and therapeutic potency
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PD-1 -cis IL-2R agonism yields better effectors from stem-like CD8 + T cells
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Targeting Cardiac Fibrosis with Engineered T cells
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A canonical to non-canonical Wnt signalling switch in haematopoietic stem-cell ageing
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FBXO38 mediates PD-1 ubiquitination and regulates anti-tumour immunity of T cells.
Nature | 2019 Jul 08 | PubMed ID: 30487606 | Read Article
MLLT3 governs human haematopoietic stem-cell self-renewal and engraftment
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Senolytic CAR T cells reverse senescence-associated pathologies
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A Homing System Targets Therapeutic T-cells to Brain Cancer
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Exit from dormancy provokes DNA-damage-induced attrition in haematopoietic stem cells.
Nature | 2015 May 14 | PubMed ID: 25707806 | Read Article
Reprogramming human T cell function and specificity with non-viral genome targeting
Nature | 2018 Jul 11 | PubMed ID: 29995861 | Read Article
CRISPR Screen in Regulatory T Cells Reveals Modulators of Foxp3
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Auto-aggressive CXCR6 + CD8 T cells cause liver immune pathology in NASH.
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Single-cell RNA-seq identifies a PD-1hi ILC progenitor and defines its development pathway.
Nature | 2016 Sep 29 | PubMed ID: 27749818 | Read Article
Tumor Regression in Patients With Metastatic Synovial Cell Sarcoma and Melanoma Using Genetically Engineered Lymphocytes Reactive With NY-ESO-1
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Chromosome segregation fidelity in epithelia requires tissue architecture
Cell | 2018 Sep 20 | PubMed ID: 30146160 | Read Article
Universal Chimeric Antigen Receptors for Multiplexed and Logical Control of T Cell Responses
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Engineered virus-like particles for efficient in vivo delivery of therapeutic proteins
Cell | 2022 Jan 20 | PubMed ID: 35021064 | Read Article
Enhanced safety and efficacy of protease-regulated CAR-T cell receptors
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Cone-shaped HIV-1 capsids are transported through intact nuclear pores
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