Despite substantial advances in the treatment of acute myeloid leukemia (AML), only 30% of AML patients survive more than 5 years. Therefore, new therapeutics are much needed. Here, we present novel therapeutic strategy targeting myeloid leukemia antigen, HLA-A2 restricted peptide PR1.Previously, we have conducted pre-clinical development and humanization of a T cell receptor-like monoclonal antibody (h8F4) that targets PR1/HLA-A2 and eliminates AML xenografts by antibody-dependent cellular cytotoxicity (ADCC) following repeat dosing. To improve the potency of h8F4, we have developed a bi-specific T cell-engaging antibody that targets PR1/HLA-A2 on leukemia and CD3 on neighboring T cells. Here we demonstrate successful production and purification of the h8F4 bi-specific antibody. Utilizing flow cytometry, we confirm PR1/HLA-A2 and CD3-specific binding characteristics, T cell activation in the presence of PR1/HLA-A2, and importantly AML target cell cytotoxicity after h8F4 bi-specific antibody engagement with healthy donor effector T cells. Cytotoxicity assays were performed with both AML cell lines and primary patient AML blasts serving as target cells and health donor PBMC as a source of effector T cells at an Effector: Target ratio of 2:1. Results indicate up to 50% leukemia-specific lysis with 0.2nM h8F4 bi-specific antibody after only 18 hours of incubation. In addition, in vivo data also confirms significant elimination of the leukemia cell line U937 (as evidenced by % leukemia cells detected in peripheral blood and bio-luminescence imaging) in an NSG-U937 AML xenograft mouse model when compared to control groups treated with effector cells alone. In conclusion, these studies demonstrate the therapeutic potential of a novel bi-specific antibody targeting the PR1/HLA-A2 leukemia-associated antigen. This bi-specific antibody appears to increase the potency of h8F4 with rapid elimination of AML and our studies justify potential development as a treatment option for patients with high-risk AML.
","abstract_html":"<p>Despite substantial advances in the treatment of acute myeloid leukemia (AML), only 30% of AML patients survive more than 5 years. Therefore, new therapeutics are much needed. Here, we present novel therapeutic strategy targeting myeloid leukemia antigen, HLA-A2 restricted peptide PR1.Previously, we have conducted pre-clinical development and humanization of a T cell receptor-like monoclonal antibody (h8F4) that targets PR1/HLA-A2 and eliminates AML xenografts by antibody-dependent cellular cytotoxicity (ADCC) following repeat dosing. To improve the potency of h8F4, we have developed a bi-specific T cell-engaging antibody that targets PR1/HLA-A2 on leukemia and CD3 on neighboring T cells. Here we demonstrate successful production and purification of the h8F4 bi-specific antibody. Utilizing flow cytometry, we confirm PR1/HLA-A2 and CD3-specific binding characteristics, T cell activation in the presence of PR1/HLA-A2, and importantly AML target cell cytotoxicity after h8F4 bi-specific antibody engagement with healthy donor effector T cells. Cytotoxicity assays were performed with both AML cell lines and primary patient AML blasts serving as target cells and health donor PBMC as a source of effector T cells at an Effector: Target ratio of 2:1. Results indicate up to 50% leukemia-specific lysis with 0.2nM h8F4 bi-specific antibody after only 18 hours of incubation. In addition, <em>in vivo </em>data also confirms significant elimination of the leukemia cell line U937 (as evidenced by % leukemia cells detected in peripheral blood and bio-luminescence imaging) in an NSG-U937 AML xenograft mouse model when compared to control groups treated with effector cells alone. In conclusion, these studies demonstrate the therapeutic potential of a novel bi-specific antibody targeting the PR1/HLA-A2 leukemia-associated antigen. This bi-specific antibody appears to increase the potency of h8F4 with rapid elimination of AML and our studies justify potential development as a treatment option for patients with high-risk AML.</p>","abstract_has_math":false,"creators":["Herrmann, Amanda","https://orcid.org/0000-0002-3912-5068
"],"institution":null,"degree_name":"Doctor of Philosophy (PhD)","degree_level":"Dissertation (PhD)","degree_discipline":null,"degree_department":null,"school":null,"contributors":["Jeffrey Molldrem MD","Gheath Al-Atrash DO, PhD","William Plunkett PhD"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2018,"date_issued":"2018-05-01T07:00:00Z","date_published":"2018-05-01T07:00:00Z","updated_at":"2026-07-24T05:49:41Z","subjects":["Acute Myeloid Leukemia","bi-specific antibody","cancer immunotherapy","re-directed cytotoxicity","Medicine and Health Sciences"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://digitalcommons.library.tmc.edu/utgsbs_dissertations/853","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Jeffrey Molldrem MD","Gheath Al-Atrash DO, PhD","William Plunkett PhD"]},{"key":"dc:creator","label":"Author","values":["Herrmann, Amanda","https://orcid.org/0000-0002-3912-5068
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"]},{"key":"dc:title","label":"Title","values":["Development and Characterization of A Bi-Specific Antibody Targeting Hla-A2/Pr1"]}]}],"canonical_facts":{"dc:contributor":["Jeffrey Molldrem MD","Gheath Al-Atrash DO, PhD","William Plunkett PhD"],"dc:creator":["Herrmann, Amanda","https://orcid.org/0000-0002-3912-5068
"],"dc:date.available":["2019-05-03T07:00:00Z"],"dc:description.abstract":["Despite substantial advances in the treatment of acute myeloid leukemia (AML), only 30% of AML patients survive more than 5 years. Therefore, new therapeutics are much needed. Here, we present novel therapeutic strategy targeting myeloid leukemia antigen, HLA-A2 restricted peptide PR1.Previously, we have conducted pre-clinical development and humanization of a T cell receptor-like monoclonal antibody (h8F4) that targets PR1/HLA-A2 and eliminates AML xenografts by antibody-dependent cellular cytotoxicity (ADCC) following repeat dosing. To improve the potency of h8F4, we have developed a bi-specific T cell-engaging antibody that targets PR1/HLA-A2 on leukemia and CD3 on neighboring T cells. Here we demonstrate successful production and purification of the h8F4 bi-specific antibody. Utilizing flow cytometry, we confirm PR1/HLA-A2 and CD3-specific binding characteristics, T cell activation in the presence of PR1/HLA-A2, and importantly AML target cell cytotoxicity after h8F4 bi-specific antibody engagement with healthy donor effector T cells. Cytotoxicity assays were performed with both AML cell lines and primary patient AML blasts serving as target cells and health donor PBMC as a source of effector T cells at an Effector: Target ratio of 2:1. Results indicate up to 50% leukemia-specific lysis with 0.2nM h8F4 bi-specific antibody after only 18 hours of incubation. In addition, in vivo data also confirms significant elimination of the leukemia cell line U937 (as evidenced by % leukemia cells detected in peripheral blood and bio-luminescence imaging) in an NSG-U937 AML xenograft mouse model when compared to control groups treated with effector cells alone. In conclusion, these studies demonstrate the therapeutic potential of a novel bi-specific antibody targeting the PR1/HLA-A2 leukemia-associated antigen. This bi-specific antibody appears to increase the potency of h8F4 with rapid elimination of AML and our studies justify potential development as a treatment option for patients with high-risk AML.
"],"dc:identifier":["https://digitalcommons.library.tmc.edu/utgsbs_dissertations/853"],"dc:subject":["Acute Myeloid Leukemia","bi-specific antibody","cancer immunotherapy","re-directed cytotoxicity","Medicine and Health Sciences"],"dc:title":["Development and Characterization of A Bi-Specific Antibody Targeting Hla-A2/Pr1"],"thesis:degree_level":["Dissertation (PhD)"],"thesis:degree_name":["Doctor of Philosophy (PhD)"]},"updated_at":"2026-07-24T05:49:41Z"}