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
AU - Alvarez-Vallina, Luis
N1 - Funding Information:
This study was supported by grants from Ministerio de Ciencia e Innovación (BIO2008-03233), Ministerio de Economía y Competitividad (BIO2011-22738), and Comunidad de Madrid (S-BIO-0236-2006 and S2010/BMD-2312) to L.A.-V. V.A.-C. was supported by a predoctoral fellowship (BFI07.132) from the Gobierno Vasco. D.S.-M. was supported by a Comunidad de Madrid/Fondo Social Europeo Training grant (FPI-000531). M.C. was supported by Instituto de Salud Carlos III (Contrato Rio Hortega, CM06/00055). The authors declared no conflict of interest.
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
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U2 - 10.1038/mtna.2013.19
DO - 10.1038/mtna.2013.19
M3 - Article
C2 - 23695536
AN - SCOPUS:84878881427
SN - 2162-2531
VL - 2
SP - e93
JO - Molecular Therapy - Nucleic Acids
JF - Molecular Therapy - Nucleic Acids
M1 - e93
ER -