Successful engineering of a highly potent single-chain variable-fragment (scFv) bispecific antibody to target disialoganglioside (GD2) positive tumors Journal Article
| Authors: | Cheng, M.; Santich, B. H.; Xu, H.; Ahmed, M.; Huse, M.; Cheung, N. K. V. |
| Article Title: | Successful engineering of a highly potent single-chain variable-fragment (scFv) bispecific antibody to target disialoganglioside (GD2) positive tumors |
| Abstract: | Engineering potent bispecific antibodies from single-chain variable fragments (scFv) remains difficult due to the inherent instability and insufficient binding of scFv's compared to their parental immunoglobulin format. Previously, we described a scFv-based bispecific antibody (scBA) against disialoganglioside (GD2) based on the anti-GD2 murine 5F11-scFv and the anti-CD3 huOKT3-scFv (5F11-scBA). In this study, we substituted the 5F11-scFv with the higher affinity (13-fold) hu3F8-scFv to form hu3F8-scBA. With this modification, hu3F8-scBA redirected T cells to kill GD2(+) cancer cell lines with up to 5,000-fold higher potency (femtomolar EC50) compared with 5F11-scBA (picomolar EC50) in cytotoxicity assays, even against target cells with low GD2 densities. Furthermore, hu3F8-scBA induced stronger T-cell activation than 5F11-scBA, as measured by Ca2+ flux and cytokine release. Additionally, in vivo, hu3F8-scBA suppressed tumor growth and prolonged mice survival much more effectively than 5F11-scBA, in both neuroblastoma and melanoma xenograft models. We conclude that the functional properties of scBA's can be increased substantially by relatively modest increases in antigen affinity. |
| Keywords: | immunotherapy; neuroblastoma; therapy; t-cells; affinity; stability; scfv; disialoganglioside gd2; anti-gd2 antibody; cancer; bispecific; tandem diabody; cell-engaging antibody |
| Journal Title: | OncoImmunology |
| Volume: | 5 |
| Issue: | 6 |
| ISSN: | 2162-4011 |
| Publisher: | Landes Bioscience |
| Date Published: | 2016-01-01 |
| Start Page: | e1168557 |
| Language: | English |
| ACCESSION: | WOS:000379162700043 |
| DOI: | 10.1080/2162402x.2016.1168557 |
| PROVIDER: | wos |
| PMCID: | PMC4938304 |
| PUBMED: | 27471647 |
| Notes: | Article -- e1168557 -- Source: Wos |
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