RetroNectin detection

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Schematic protocol for infecting target cells using RetroNectin

Schematic protocol for infecting target cells using RetroNectin reagent. In the RBV (RetroNectin-bound virus) method, the retrovirus is first bound to the RetroNectin-coated plate, and cells are added after removing the retrovirus supernatant. In this protocol, removal of the supernatant reduces inhibitory molecules that otherwise can reduce the transduction efficiency.

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RetroNectin reagent is a recombinant human fibronectin fragment that contains three functional domains: the cell-binding domain (C-domain), the heparin-binding domain (H-domain), and the CS-1 sequence

RetroNectin reagent is a recombinant human fibronectin fragment that contains three functional domains: the cell-binding domain (C-domain), the heparin-binding domain (H-domain), and the CS-1 sequence. RetroNectin Reagent enhances lentiviral- and retroviral-mediated gene transduction by aiding the colocalization of target cells and viral particles. Specifically, virus particles bind RetroNectin Reagent via interaction with the H-domain, and target cells bind mainly through the interaction of cell surface integrin receptors VLA-5 and/or VLA-4 with the fibronectin C-domain and CS-1 sites, respectively. By facilitating close physical proximity, RetroNectin Reagent can enhance viral-mediated gene transfer to target cells expressing integrin receptors VLA-4 and/or VLA-5.

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Comparison of T cell expansion efficiencies for various commonly used protocols

Comparison of T cell expansion efficiencies for various commonly used protocols. Cells co-stimulated using RetroNectin and α-CD3 demonstrated higher-fold expansion as compared to other protocols (Panel A). In addition, the presence of RetroNectin promoted the production of more naïve-like T cells (CCR7+/CD45RA+ phenotype) that have been shown to have better in vivo anti-tumor activity (Panel B).

NOTE: This expansion method requires a license for commercial applications.

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Comparison of retroviral transduction of a K562 cell line using transduction enhancers from different companies

Comparison of retroviral transduction of a K562 cell line using transduction enhancers from different companies. A K562 cell line was transduced with a retroviral vector in the presence of various transduction enhancers. RetroNectin yielded a far higher transduction efficiency relative to each of the other transduction enhancers, including Polybrene, protamine sulfate, and human fibronectin from three other companies.

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Efficient transduction of human PBMCs using RetroNectin, as compared to Polybrene

Efficient transduction of human PBMCs using RetroNectin, as compared to Polybrene. Human PBMCs were transduced with varying dilutions of lentivirus encoding ZsGreen1 in the presence of RetroNectin or Polybrene, and the percentages of transduced CD8+ cells were measured by flow cytometry. Transduction efficiencies with Retronectin were significantly higher than those obtained using Polybrene, especially at low MOI (90X dilution).

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T-cell expansion using RetroNectin reagent.

T-cell expansion using RetroNectin reagent. Peripheral blood T cells were stimulated and expanded in vitro using anti-CD3 (clone OKT3) and IL-2 alone or in combination with RetroNectin reagent (RN), anti-CD28, or anti-4-1BB. Stimulation with RetroNectin reagent yielded higher proportions of cytotoxic CD8+ T cells.