The most attractive feature of exosomes is that exosomes contain proteins, RNAs and lipids. The surface components of exosomes will decide the tissue-specific uptake of exosomes, while the inside contents of exosomes, such as RNAs, proteins, which can be engineered, will module the behavior of target cells. Therefore, exosome could be an attractive delivery vesicle for RNAs, proteins and small molecule compounds. Evomic Science developed unique proprietary techniques to load your desired proteins and /or small RNAs and/or hydrophobic compounds into exosomes.
1. Protein loading into exosome.
1.1. Regarding to exosome from cell culture. We developed a unique proprietary technique, ExoSort Technology, to target any peptide or protein into exosome. This technology is based on exosome protein sorting pathways, such as ESCRT dependent- and independent- pathways. When the desired peptides or proteins are tagged with exosome sorting motifs, the peptides or proteins will be sorted into the exosomes during exosome formation. Depending on the protein features, soluble proteins could be packed to the surface or inside of exosomes (Fig.1A), while membrane-bound proteins could be inserted into exosome membrane (Fig.1B). This sorting process of peptides or proteins, through a natural exosome sorting pathway, ensures the correct folding, posttranslational modification, and proper destination of your desired proteins or peptides. Therefore, the proteins loaded into exosomes could be used for the development of the monoclonal antibody, the efficient vaccine development, tumor immunotherapy, molecular diagnostics, and even drug screening.
We have successfully sorted viral proteins, GPCRs, transporters and Pan Fc Receptors as well as reporter proteins such as luciferase, toxic proteins and red fluorescent proteins (RFP) into exosomes in cell lines. Those engineering exosomes could be used as internal/spiked exosome control/standard for evaluating exosome isolation efficiency; tracking exosome fate by monitoring real-time exosomes both in vitro and in vivo; viral antibody and auto-antibody detection in patients; antibody-directed tissue-specific delivery of exosomes; and other animal model study.
1.2, Regarding to exosomes from body fluids, such as serum, plasma, and urine etc, we have the luciferase fused exosome proteins, which could specifically bound to exosomes. After incubating exosomes with the reporter proteins, your exosomes would be labeled with luciferase. The unloaded reporter protein could be removed using our exosome purification column kit.
2. siRNA or miRNA Loading into Exosome.
small RNAs can be loaded into exosomes by 1). extracellular loading after exosome isolation or 2). intracellular sorting during exosome formation. Intracellular loading of small RNAs is based on the 3'UTR motifs of miRNA. The most efficient extracellular loading method of siRNA or miRNA is electroporation. We developed a Exosome Electroporation Buffer which can efficiently load small RNA into exosomes, maintain the integration of exosomes, and reduce the exosome aggregation caused by electroporation. After electroporation, unloaded siRNA or miRNA can be removed by our exosome purification column kit (ExoA300-B300-C300). The purified exosomes would be used for downstream application.
3. Compound loading into exosome.
Delivery of hydrophobic compound into cells in vitro and in vivo has been a challenge task. We developed a featured Compound Loading Buffer, which can efficiently encapsulate small hydrophobic compounds into exosomes under electroporation. Free compounds will be removed from exosome mixture by exosome column purification kit.
Fig.3. Sorting your target proteins into exosome using ExoSort Technology. A speculative model of an exosome with lipid bilayer membrane is showed. Soluble proteins, tagged with exosome sorting motifs, could be sorted into inside of exosomes (Left panel). Callouts, outlined with black, red, or green color, could be your target proteins. Membrane-bound proteins, tagged with exosome sorting motifs, could be inserted into the exosome membrane (Right panel).
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