{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T08:45:41Z","timestamp":1792140341408,"version":"4.4.0"},"reference-count":62,"publisher":"American Chemical Society (ACS)","issue":"10","license":[{"start":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T00:00:00Z","timestamp":1790467200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000054","name":"National Cancer Institute","doi-asserted-by":"publisher","award":["1DP2CA280622"],"award-info":[{"award-number":["1DP2CA280622"]}],"id":[{"id":"10.13039\/100000054","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100023015","name":"Advanced Research Projects Agency for Health","doi-asserted-by":"publisher","award":["AY2AX0000006"],"award-info":[{"award-number":["AY2AX0000006"]}],"id":[{"id":"10.13039\/100023015","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,10,16]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Bispecific T cell engagers (BiTEs) are engineered immunotherapeutic molecules designed to direct the body\u2019s immune system against malignant cells by physically connecting T cells to target cells, triggering cytotoxic responses. BiTEs have typically engaged T cells via \u03b1CD3 domains, such as OKT3 or UCHT1, which are linked to a second binding domain specific for a tumor-associated antigen. The risk of on-target off-tumor toxicity is a serious concern for BiTE immunotherapy due to the expression of tumor-associated antigen targets on healthy tissues. To address this concern, we have developed protease-responsive masked-BiTEs using OKT3 and UCHT1 mAb clones, that activate T cells only after exposure to tumor micro-environment-associated proteases and stimulate the T cell activation by their tumor-antigenic targets. Our protease-responsive masked BiTEs (mBiTEs) are comprised of \u03b1CD3 Fab proteins that are sterically prevented from binding CD3 via N\u2019 terminal fusion of coiled-coil leucine zipper dimerization domains to each Fab chain, with further C\u2019 terminal linkage to a tumoral-antigen targeting nanobody (either \u03b1PSMA or \u03b1HER2). The peptide linking each Fab chain and its coiled-coil masking zipper harbors a protease cleavable site, such that engagement of either recombinant or T cell displayed CD3 is significantly reduced until mBiTE protease activation. OKT3 and UCHT1-based masked T cell engagers (mTEs) showed \u223c400-fold and \u223c800-fold reduced affinity for CD3 protein compared to unmodified T cell engagers (TEs), with \u223c222-fold and \u223c457-fold restoration of binding upon proteolytic unmasking, respectively. mBiTEs did not activate Jurkat T cells while proteolytically unmasked proteins did, and mBiTEs harboring linkers cleavable by the cancer associated matric metalloprotease 2 (MMP2) enzyme enabled killing of PSMA + and HER2 + cancer cells in an MMP2-dependent manner in PBMC co-culture assays. The mBiTEs we describe here could be of broad utility towards helping constrain BiTE-mediated immune-activation to protease-high environments, such as the solid tumor microenvironment.<\/jats:p>","DOI":"10.1021\/acschembio.6c00688","type":"journal-article","created":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T19:01:33Z","timestamp":1790535693000},"page":"2774-2786","source":"Crossref","is-referenced-by-count":0,"title":["Proteolytic Activation of T Cell Engagers And Bispecific Proteins Using Modular Coiled-Coil Peptides"],"prefix":"10.1021","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6676-9846","authenticated-orcid":true,"given":"Basudeb","family":"Mondal","sequence":"first","affiliation":[{"name":"Georgia Institute of Technology , , , ,","place":["Atlanta, Georgia, United States, 30332"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yusef A","family":"Haikal","sequence":"additional","affiliation":[{"name":"Georgia Institute of Technology , , , ,","place":["Atlanta, Georgia, United States, 30332"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6255-6755","authenticated-orcid":true,"given":"Gabriel A.","family":"Kwong","sequence":"additional","affiliation":[{"name":"Emory University and Georgia Institute of Technology , , , ,","place":["Atlanta, Georgia, United States, 30332"]},{"name":"Georgia Institute of Technology , , , ,","place":["Atlanta, Georgia, United States, 30332"]},{"name":"Georgia Institute of Technology , , , ,","place":["Atlanta, Georgia, United States, 30332"]},{"name":"Emory University and Georgia Institute of Technology , , , ,","place":["Atlanta, Georgia, United States, 30332"]},{"name":"Winship Cancer Institute of Emory University , , ,","place":["Atlanta, Georgia, United States, 30322"]},{"name":"Georgia Institute of Technology , , , ,","place":["Atlanta, Georgia, United States, 30332"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6110-2938","authenticated-orcid":true,"given":"John","family":"Blazeck","sequence":"additional","affiliation":[{"name":"Georgia Institute of Technology , , , ,","place":["Atlanta, Georgia, United States, 30332"]},{"name":"Georgia Institute of Technology , , , ,","place":["Atlanta, Georgia, United States, 30332"]},{"name":"Georgia Institute of Technology , , , ,","place":["Atlanta, Georgia, United States, 30332"]},{"name":"Emory University and Georgia Institute of Technology , , , ,","place":["Atlanta, Georgia, United States, 30332"]},{"name":"Winship Cancer Institute of Emory University , , ,","place":["Atlanta, Georgia, United States, 30322"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,27]]},"reference":[{"issue":"3","key":"2026101523065568000_cit1","doi-asserted-by":"publisher","first-page":"133","DOI":"10.1038\/s41568-019-0116-x","article-title":"The evolving landscape of biomarkers for checkpoint inhibitor immunotherapy","volume":"19","author":"Havel","year":"2019","journal-title":"Nat. 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