BioCat GmbH • Im Neuenheimer Feld 584 • D-69120 Heidelberg • Tel.: +49(0)6221 7141516
Home -> Cell Biology -> Exosomes -> Exosome Targeting

Exosome Targeting

• Display proteins on exosome surfaces
• Coat exosomes with targeting ligands
• Target specific cellular destinations
• Create stable XStamp cell lines

XStamp™ Exosome Targeting Technology

Program Exosomes with Specific Addresses
Exosomes are extracellular nanoshuttles that facilitate communication between cells and can be engineered as therapeutic shuttles to deliver biological molecules or drugs to target disease cells. SBI has developed an exosome surface display system that enables desired protein sequences to be placed efficiently on the surfaces of engineered exosomes called the "XStamp" technology. The patented XStamp technology is based upon a C-terminal fusion of the C1C2 domain from MFG-E8. Protein sequences that are fused to the XStamp tag will efficiently display the protein ligand fusion on the surfaces of secreted exosomes. The technology can be used to place cellular "addresses" on exosomes that send them to specific destinations for cargo delivery.

MFG-E8 Localizes to Exosome Surfaces (C1C2 domain)

Fluorescently-labeled antibodies for MFG-E8, CD58 and CD81 were used in combination for FACs analysis. The CD58 marker is a known cell surface marker that is absent on exosomes. CD81 is known to be present both in cells and exosomes. The FACs data show that MFG-E8 is exclusively detected on exosomes and not present in the cells.

The XStamp System
To take advantage of the localization of MFG-E8 on exosomes, the C1C2 domain (XStamp domain) of the protein´s gene was cloned into SBI´s MSCV-MCS-EF1-Puro lentivector and a 5´ secretion signal sequence (SS) was placed within the multiple cloning site.
The protein ligand chosen to display on exosomes is cloned into the MCS and fused to the C1C2 domain. The XStamp lentivector also features a downstream EF1-Puromycin cassette for selection and stable cell line development. The lentivector constructs can be used in transient transfection expression studies as well as for packaging into lentivirus to stably transduce cells to create cellular factories producing engineered exosomes.

Motilin-XStamp for GI tract targeting
Motilin is a 22-amino acid polypeptide hormone that binds to the Motilin receptor which is exclusively expressed in the intestine. To test the Motilin-XStamp construct (catalog# XSTP720PA-1-SBI), it was transfected into HEK293 cells and after 48 hours, the exosomes were collected using ExoQuick-TC. The next day, the XStamp-Motilin exosomes were Exo-Fected with a Texas-Red-labeled siRNA to monitor exosome docking and delivery. The transfected XStamp-Motilin exosomes were then added to MDA-MB-231 Breast Cancer Cells (motilin receptor negative) and to HT-29 Colon Cancer Cells (motilin receptor positive).

The cells were imaged after 24 hours for uptake of the Texas-Red-labeled siRNA delivery from the XStamped exosomes. The HT-29 colon cancer cells that are motilin receptor positive took up the XStamp-Motilin exosomes at a much higher rate than the MDA-MB-231 Breast Cancer (motilin receptor negative) cells.

Targeting Neurons using XStamps and Exosomes
An NCAM-XStamp fusion was constructed which incorporated the first 300 amino acids (Signal peptide plus IGc2 domains 1-3) of the mouse NCAM gene translationally fused to the C1C2 XStamp display tag (catalog# XSTP721PA-1). In parallel, a Brain Homing Peptide (BHP1, from "Organ targeting in vivo using phage display peptide libraries." Pasqualini R, et al. Nature (6572):364-6) was fused to the C1C2 XStamp domain to create the BHP1-XStamp construct (catalog# XSTP722PA-1-SBI).


NCAM-XStamp and Brain Homing Peptide 1 for Neural Targeting
The NCAM-XStamp construct (catalog# XSTP721PA-1-SBI) and the BHP1-XStamp construct (catalog# XSTP722PA-1-SBI), were transfected separately into mouse MSCs along with SBI´s XPack-GFP construct (catalog# XPAK530PA-1) which packages GFP into the interior of exosomes for fluorescent tracking. After 48 hours, the exosomes were collected. Equal amounts (100 ug) of Control (No XStamp), NCAM-XStamp or BHP1-XStamp (Brain Homing Peptide) MSC exosomes (all loaded with XPack-GFP) were added to Neuro2a neuroblastoma cells in culture. The neurons were imaged for phase and GFP signals after a 24 hour incubation with the various exosomes to monitor GFP delivery mediated by the XStamp ligand coats. Taken together, this is known as the "Pack and Stamp" system, which can be adapted for any cargo and any delivery target.


Technical References

Batrakova EV, Kim MS. Using exosomes, naturally-equipped nanocarriers, for drug delivery. J Control Release. 2015 Aug 1. pii: S0168-3659(15)30042-0.

Ohno S, Takanashi M, Sudo K, Ueda S, Ishikawa A, Matsuyama N, Fujita K, Mizutani T, Ohgi T, Ochiya T, Gotoh N, Kuroda M. Systemically injected exosomes targeted to EGFR deliver antitumor microRNA to breast cancer cells. Mol Ther. 2013 Jan;21(1):185-91.

Delcayre A, Le Pecq JB. Exosomes as novel therapeutic nanodevices. Curr Opin Mol Ther. 2006 Feb;8(1):31-8. Review.

Hartman ZC, Wei J, Glass OK, Guo H, Lei G, Yang XY, Osada T, Hobeika A, Delcayre A, Le Pecq JB, Morse MA, Clay TM, Lyerly HK. Increasing vaccine potency through exosome antigen targeting. Vaccine. 2011 Nov 21;29(50):9361-7.

Cheng L, Huang FZ, Cheng LF, Zhu YQ, Hu Q, Li L, Wei L, Chen DW. GE11-modified liposomes for non-small cell lung cancer targeting: preparation, ex vitro and in vivo evaluation. Int J Nanomedicine. 2014 Feb 12;9:921-35.

Pasqualini R, Ruoslahti E. Organ targeting in vivo using phage display peptide libraries. Nature. 1996 Mar 28;380(6572):364-6.

Estelles A, Sperinde J, Roulon T, Aguilar B, Bonner C, LePecq JB, Delcayre A. Exosome nanovesicles displaying G protein-coupled receptors for drug discovery. Int J Nanomedicine. 2007;2(4):751-60.

Pirjo Laakkonen and Kirsi Vuorinena. Homing peptides as targeted delivery vehicles. Integr. Biol., 2010,2, 326-337.

Morse MA, Garst J, Osada T, Khan S, Hobeika A, Clay TM, Valente N, Shreeniwas R, Sutton MA, Delcayre A, Hsu DH, Le Pecq JB, Lyerly HK. A phase I study of dexosome immunotherapy in patients with advanced non-small cell lung cancer. J Transl Med. 2005 Feb 21;3(1):9.

Rountree RB, et al. Exosome targeting of tumor antigens expressed by cancer vaccines can improve antigen immunogenicity and therapeutic efficacy.Cancer Res. 2011 Aug 1;71(15):5235-44.

van der Meel R, Fens MH, Vader P, van Solinge WW, Eniola-Adefeso O, Schiffelers RM. Extracellular vesicles as drug delivery systems: lessons from the liposome field. J Control Release. 2014 Dec 10;195:72-85.

Gyorgy B, Hung ME, Breakefield XO, Leonard JN. Therapeutic applications of extracellular vesicles: clinical promise and open questions. Annu Rev Pharmacol Toxicol. 2015;55:439-64.
PDF-Downloads - Will open in new browser window

Brochure Exosome - Research, Products and Services



Description Cat# Size Price    
XStamp cloning and expression lentivector MSCV-Leader-MCS-C1C2-EF1-Puro XSTP710PA-1-SBI 10 ug 802 € DETAILS   Add to Cart 
XStamp BHP1 exosome brain homing peptide targeting lentivector XSTP722PA-1-SBI 10 ug 830 € DETAILS   Add to Cart 
XStamp CD40L exosome CD40 receptor targeting lentivector XSTP725PA-1-SBI 10 ug 830 € DETAILS   Add to Cart 
XStamp GE11 exosome EGFR targeting lentivector XSTP723PA-1-SBI 10 ug 830 € DETAILS   Add to Cart 
XStamp Her2 ligand exosome Her2 receptor targeting lentivector XSTP724PA-1-SBI 10 ug 830 € DETAILS   Add to Cart 
XStamp IL2 exosome IL-2 receptor targeting lentivector XSTP726PA-1-SBI 10 ug 830 € DETAILS   Add to Cart 
XStamp mlN exosome Motilin receptor GI tract targeting lentivector XSTP720PA-1-SBI 10 ug 830 € DETAILS   Add to Cart 
XStamp NCAM exosome brain targeting lentivector XSTP721PA-1-SBI 10 ug 830 € DETAILS   Add to Cart 

BioCat Summer Special

Summer Special

10% OFF
Norgen Sample Preparation Kits


30% OFF BioCat Universal Agarose

Email Newsletter

Subscribe to the BioCat Email Newsletter.


Logos Biosystems Cell Counters

Cell-based Assays

Cell-based assays

Choose from a wide variety of cell-based assays designed for greater accuracy and increased throughput.

50% OFF Fluorescent Proteins

Fluorescent Protein Vectors

Get 50% discount on the second and subsequent vectors when ordering two or more vectors with the same fluorescent protein.

Lentiviral Barcode Library

Lentiviral-based barcode library

Track clonal populations of cells over time or through different treatments using the lentiviral-based barcode library.

Imprint / Impressum | Privacy Policy / Datenschutzerklärung
Top of Page Up!