| RRC ID |
57860
|
| Author |
Kurosawa N, Wakata Y, Ida K, Midorikawa A, Isobe M.
|
| Title |
High throughput development of TCR-mimic antibody that targets survivin-2B80-88/HLA-A*A24 and its application in a bispecific T-cell engager.
|
| Journal |
Sci Rep
|
| Abstract |
Intracellular tumor-associated antigens are targeted by antibodies known as T-cell receptor mimic antibodies (TCRm-Abs), which recognize T-cell epitopes with better stabilities and higher affinities than T-cell receptors. However, TCRm-Abs have been proven difficult to produce using conventional techniques. Here, we developed TCRm-Abs that recognize the survivin-2B-derived nonamer peptide, AYACNTSTL (SV2B80-88), presented on HLA-A*24 (SV2B80-88/HLA-A*24) from immunized mice by using a fluorescence-activated cell sorting-based antigen-specific plasma cells isolation method combined with a high-throughput single-cell-based immunoglobulin-gene-cloning technology. This approach yielded a remarkable efficiency in generating candidate antibody clones that recognize SV2B80-88/HLA-A*24. The screening of the antibody clones for their affinity and ability to bind key amino-acid residues within the target peptide revealed that one clone, #21-3, specifically recognized SV2B80-88/HLA-A*24 on T2 cells. The specificity of #21-3 was further established through survivin-2B-positive tumor cell lines that exogenously or endogenously express HLA-A*24. A bispecific T-cell engager comprised of #21-3 and anti-CD3 showed specific cytotoxicity towards cells bearing SV2B80-88/HLA-A*24 by recruiting and activating T-cells in vitro. The efficient development of TCRm-Ab overcomes the limitations that hamper antibody-based immunotherapeutic approaches and enables the targeting of intracellular tumor-associated antigens.
|
| Volume |
9(1)
|
| Pages |
9827
|
| Published |
2019-7-8
|
| DOI |
10.1038/s41598-019-46198-5
|
| PII |
10.1038/s41598-019-46198-5
|
| PMID |
31285464
|
| PMC |
PMC6614450
|
| MeSH |
Animals
Antibodies, Bispecific / metabolism*
Cell Separation
Epitopes, T-Lymphocyte / immunology
Flow Cytometry
HLA-A24 Antigen / immunology*
HeLa Cells
High-Throughput Screening Assays
Humans
Immunization
Leukocytes, Mononuclear / cytology
Mice
Receptors, Antigen, T-Cell / immunology*
Survivin / immunology*
|
| IF |
3.998
|
| Times Cited |
3
|
| Resource |
| Human and Animal Cells |
HeLa(RCB0007)
Jurkat(RCB3052)
COLO-320(RCB1193) |
