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Products >  Molecular_Biology >  Gene_Transfer_and_Expression >  RetroNectin_Reagent_and_Precoated_Dishes

Research and Clinical Grade RetroNectin Reagent for Retroviral or Lentiviral Transduction

RetroNectin Reagent for retroviral or lentiviral transduction significantly increases transduction efficiency and results in enhanced gene transfer, especially in suspension and hematopoietic stem cells. RetroNectin Reagent is a chimeric peptide composed of E. coli-derived recombinant human fibronectin fragment that contains three functional domains: a central cell-binding domain (type III repeat, 8-10), a heparin-binding domain II (type III repeat, 12-14), and a CS1 site within the alternatively-spliced IIICS region. RetroNectin is a 574 amino acid protein with a molecular weight of 63 kDa. Fibronectins (FNS) are multi-functional cell adhesive glycoproteins present in the extracellular cell matrix and plasma and also play critical roles in wound healing and embryogenesis.

When coated on surfaces of cell culture dishes, flasks or bags, RetroNectin significantly increases retroviral or lentiviral transduction into mammalian cells. This enhanced gene transfer is thought to occur via the co-localization of retroviral particles and target cells on the RetroNectin molecules. In essence, viral particles bind RetroNectin via interaction with its heparin-binding domain II, while target cells bind RetroNectin primarily through interaction of the cell surface integrin receptor VLA-4 with the fibronectin CS1 site. Additionally, target cells may also bind the RetroNectin recombinant human fibronectin fragment via the interaction of fibronectin tripeptide Arg-Gly-Asp (RGD), located within the molecule's central cell-binding domain, with corresponding cell surface integrin receptor VLA-5.


Improve Viral Transductions with RetroNectin Reagent

Recombinant human fibronectin fragment colocalizes virus and cells to facilitate enhanced gene transfer

    • Improves gene transfer efficiency in hard-to-infect cell types and hematopoietic stem cells
    • Multivalent molecule simultaneously binds virus particles and cell surface proteins, maximizing cell-virus contact
    • Ideal for use with Lenti-X and Retro-X Expression Systems

  At-A-Glance   Documents   Images & Data   Resources

Features

  • Enhanced gene transfer efficiency
  • Higher retroviral or lentiviral transduction levels in suspended and adherent mammalian cells
  • Improved success in hard-to-infect cell types and hematopoietic stem cells
  • Nontoxic: does not require cytotoxic polybrene
  • Superior transduction efficiency compared with fibronectin, polybrene or protamine
  • Higher reagent efficiency enables use of smaller viral solution volumes
  • Clinical grade RetroNectin is available (for GMP grade RetroNectin only please complete and return this form prior to purchase)

Applications

  • General retroviral or lentiviral transduction experiments
  • Cell migration assays
  • Gene therapy studies: RetroNectin has already been used in over 40 clinical trials, mostly using hematopoietic stem cells
  • Cell therapy studies related to stem cell research and related technology

Form

  • Research Grade RetroNectin (Cat. # T100A/B): Sterilized solution in 12.5 mM sodium citrate (pH 6.2) and 1.25% sucrose.
  • Clinical Grade RetroNectin (Cat. # T201): 2.5 mg lyophilized powder
  • RetroNectin Dishes (Cat. # T110A): 10 precoated 3.5 cm dishes

Notes

  • Research Grade RetroNectin (Cat. # T100A/B; T110A): For research use only. Not for use in diagnostic or therapeutic procedures.
  • Clinical Grade RetroNectin (GMP) (Cat. # T201): RetroNectin (GMP) is manufactured as a quality-assured product, according to guidelines for Good Manufacturing Practice (GMP) for Investigational Products. This product (Cat. #T201) can be used for ex vivo clinical applications and is limited to investigational use only.

Plate Coating

It is recommended that RetroNectin Reagent be coated onto plates at a concentration of 20–100 µg/mL (4–20 µg/cm2). RetroNectin Precoated Dishes are precoated (40 µg/mL) 3.5 cm diameter dishes and are ready to use.

Cell Transduction Efficiencies using RetroNectin
Cell Line/TypeEfficiency of Gene Transfer (%)
TF-153.3
HEL28.1
NIH/3Y380.5
CD34+ bMC 86.9
Mice BM MNC fraction 29.2
K-56293.5
HL-6089.4

References

  1. Asada, K. et al. (1998) J. Biochem. 123:1041-1047.
  2. Chono, H. et al. (2001) J. Biochem. 130:331-334.
  3. Chono, H. et al. (2011) J. Biochem. 149:285-292.
  4. Fukushima, K. et al. (2009) J. Biol. Chem. 284:7719-7732.
  5. Hanenberg, H. et al. (1996) Nature Medicine 2:876-882.
  6. Hiasa, A. et al. (2009) Gene Therapy 16:620-628.
  7. Kawahara, M. et al. (2008) Blood 11(9):3856-3866.
  8. Kimizuka, F. et al. (1991) J. Biochem. 110:284-291.
  9. Okamoto, S. et al. (2009) Cancer Res. 69:9003-9011.
  10. Yu, S.S. et al. (2008) Cancer Gene Therapy 15:508-516.



 
 
Products
Cat. # Product Contents Size Price License Units Select
T201 RetroNectin® (GMP) 2.5 mg $1,400.00 License Statements
T110A RetroNectin® Precoated Dish 10 Dishes $331.00 License Statements
T100B RetroNectin® Recombinant Human Fibronectin Fragment 2.5 mg $645.00 License Statements
T100A RetroNectin® Recombinant Human Fibronectin Fragment 0.5 mg $166.00 License Statements
 

 

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