Abstract
A human single-chain variable fragment (scFv) antibody library was expressed on the surface of human T cells after transduction with lentiviral vectors (LVs). The repertoire was fused to a first-generation T cell receptor ζ (TCRζ)-based chimeric antigen receptor (CAR). We used this library to isolate antibodies termed CARbodies that recognize antigens expressed on the tumor cell surface in a proof-of-principle system. After three rounds of activation-selection there was a clear repertoire restriction, with the emergence dominant clones. The CARbodies were purified from bacterial cultures as soluble and active proteins. Furthermore, to validate its potential application for adoptive cell therapy, human T cells were transduced with a LV encoding a second-generation costimulatory CAR (CARv2) bearing the selected CARbodies. Transduced human primary T cells expressed significant levels of the CARbodies-based CARv2 fusion protein on the cell surface, and importantly could be specifically activated, after stimulation with tumor cells. This approach is a promising tool for the generation of antibodies fully adapted to the display format (CAR) and the selection context (cell synapse), which could extend the scope of current adoptive cell therapy strategies with CAR-redirected T cells.
| Original language | English (US) |
|---|---|
| Article number | e93 |
| Journal | Molecular Therapy - Nucleic Acids |
| Volume | 2 |
| DOIs | |
| State | Published - 2013 |
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Keywords
- Adoptive cell therapy
- Antibody
- Chimeric antigen receptor
- Repertoire selection
- Tumor-associated antigens
ASJC Scopus subject areas
- Molecular Medicine
- Drug Discovery
Cite this
CARbodies : Human antibodies against cell surface tumor antigens selected from repertoires displayed on T cell chimeric antigen receptors. / Alonso-Camino, Vanesa; Sánchez-Martín, David; Compte, Marta; Nuñez-Prado, Natalia; Diaz, Rosa M.; Vile, Richard Geoffrey; Alvarez-Vallina, Luis.
In: Molecular Therapy - Nucleic Acids, Vol. 2, e93, 2013.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - CARbodies
T2 - Human antibodies against cell surface tumor antigens selected from repertoires displayed on T cell chimeric antigen receptors
AU - Alonso-Camino, Vanesa
AU - Sánchez-Martín, David
AU - Compte, Marta
AU - Nuñez-Prado, Natalia
AU - Diaz, Rosa M.
AU - Vile, Richard Geoffrey
AU - Alvarez-Vallina, Luis
PY - 2013
Y1 - 2013
N2 - A human single-chain variable fragment (scFv) antibody library was expressed on the surface of human T cells after transduction with lentiviral vectors (LVs). The repertoire was fused to a first-generation T cell receptor ζ (TCRζ)-based chimeric antigen receptor (CAR). We used this library to isolate antibodies termed CARbodies that recognize antigens expressed on the tumor cell surface in a proof-of-principle system. After three rounds of activation-selection there was a clear repertoire restriction, with the emergence dominant clones. The CARbodies were purified from bacterial cultures as soluble and active proteins. Furthermore, to validate its potential application for adoptive cell therapy, human T cells were transduced with a LV encoding a second-generation costimulatory CAR (CARv2) bearing the selected CARbodies. Transduced human primary T cells expressed significant levels of the CARbodies-based CARv2 fusion protein on the cell surface, and importantly could be specifically activated, after stimulation with tumor cells. This approach is a promising tool for the generation of antibodies fully adapted to the display format (CAR) and the selection context (cell synapse), which could extend the scope of current adoptive cell therapy strategies with CAR-redirected T cells.
AB - A human single-chain variable fragment (scFv) antibody library was expressed on the surface of human T cells after transduction with lentiviral vectors (LVs). The repertoire was fused to a first-generation T cell receptor ζ (TCRζ)-based chimeric antigen receptor (CAR). We used this library to isolate antibodies termed CARbodies that recognize antigens expressed on the tumor cell surface in a proof-of-principle system. After three rounds of activation-selection there was a clear repertoire restriction, with the emergence dominant clones. The CARbodies were purified from bacterial cultures as soluble and active proteins. Furthermore, to validate its potential application for adoptive cell therapy, human T cells were transduced with a LV encoding a second-generation costimulatory CAR (CARv2) bearing the selected CARbodies. Transduced human primary T cells expressed significant levels of the CARbodies-based CARv2 fusion protein on the cell surface, and importantly could be specifically activated, after stimulation with tumor cells. This approach is a promising tool for the generation of antibodies fully adapted to the display format (CAR) and the selection context (cell synapse), which could extend the scope of current adoptive cell therapy strategies with CAR-redirected T cells.
KW - Adoptive cell therapy
KW - Antibody
KW - Chimeric antigen receptor
KW - Repertoire selection
KW - Tumor-associated antigens
UR - http://www.scopus.com/inward/record.url?scp=84878881427&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84878881427&partnerID=8YFLogxK
U2 - 10.1038/mtna.2013.19
DO - 10.1038/mtna.2013.19
M3 - Article
C2 - 23695536
AN - SCOPUS:84878881427
VL - 2
JO - Molecular Therapy - Nucleic Acids
JF - Molecular Therapy - Nucleic Acids
SN - 2162-2531
M1 - e93
ER -