{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T04:49:13Z","timestamp":1790830153099,"version":"4.1.0"},"reference-count":45,"publisher":"American Society for Microbiology","issue":"7","license":[{"start":{"date-parts":[[2024,7,17]],"date-time":"2024-07-17T00:00:00Z","timestamp":1721174400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2024,7,17]],"date-time":"2024-07-17T00:00:00Z","timestamp":1721174400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"name":"HHS | National Institutes of Health","award":["R01AI125462"],"award-info":[{"award-number":["R01AI125462"]}]},{"name":"HHS | National Institutes of Health","award":["75N93019C00051"],"award-info":[{"award-number":["75N93019C00051"]}]},{"name":"HHS | National Institutes of Health","award":["T32-AI070117"],"award-info":[{"award-number":["T32-AI070117"]}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["mBio"],"published-print":{"date-parts":[[2024,7,17]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      Monoclonal antibodies (mAbs) are an attractive therapeutic platform for the prevention and treatment of influenza virus infection. There are two major glycoproteins on the influenza virion surface: hemagglutinin (HA), which is responsible for viral attachment and entry, and neuraminidase (NA), which mediates viral egress by enzymatically cleaving sialic acid to release budding particles from the host cell surface. Broadly neutralizing antibodies (bNAbs) that target the conserved HA central stalk region, such as CR9114, can inhibit both viral entry and egress. More recently, broadly binding mAbs that engage and inhibit the NA active site, such as 1G01, have been described to prevent viral egress. Here, we engineered bispecific antibodies (bsAbs) that combine the variable domains of CR9114 and 1G01 into a single molecule and evaluated if simultaneous targeting of two different glycoproteins improved antiviral properties\n                      <jats:italic>in vitro<\/jats:italic>\n                      and\n                      <jats:italic>in vivo<\/jats:italic>\n                      . Several CR9114\/1G01 bsAbs were generated with various configurations of the two sets of the variable domains (\u201cbsAb formats\u201d). We found that combinations employing the addition of a single-chain variable fragment in the hinge region of an IgG scaffold had the best properties in terms of expression, stability, and binding. Further characterization of selected bsAbs showed potent neutralizing and egress-inhibiting activity. One such bsAb (\u201chSC_CR9114_1G01\u201d) provided higher levels of prophylactic protection from mortality and morbidity upon challenge with H1N1 than either of the parental mAbs at low dosing (1 mg\/kg). These results highlight the potential use of bsAbs that simultaneously target HA and NA as new influenza immunotherapeutics.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>Infection by the influenza virus remains a global health burden. The approaches utilized here to augment the activity of broadly protective influenza virus antibodies may lead to a new class of immunotherapies with enhanced activity.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/mbio.01085-24","type":"journal-article","created":{"date-parts":[[2024,6,20]],"date-time":"2024-06-20T09:01:37Z","timestamp":1718874097000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":9,"title":["Broadly protective bispecific antibodies that simultaneously target influenza virus hemagglutinin and neuraminidase"],"prefix":"10.1128","volume":"15","author":[{"given":"Kevin E.","family":"Ramos","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Albert Einstein College of Medicine, Bronx, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nisreen M. A.","family":"Okba","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, USA"},{"name":"Center for Vaccine Research and Pandemic Preparedness (C-VaRPP), Icahn School of Medicine at Mount Sinai, New York, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica","family":"Tan","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pooja","family":"Bandawane","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, USA"},{"name":"Center for Vaccine Research and Pandemic Preparedness (C-VaRPP), Icahn School of Medicine at Mount Sinai, New York, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Philip S.","family":"Meade","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Madhumathi","family":"Loganathan","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Benjamin","family":"Francis","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sergey","family":"Shulenin","sequence":"additional","affiliation":[{"name":"Integrated BioTherapeutics, Inc., Rockville, Maryland, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Frederick W.","family":"Holtsberg","sequence":"additional","affiliation":[{"name":"Integrated BioTherapeutics, Inc., Rockville, Maryland, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M. Javad","family":"Aman","sequence":"additional","affiliation":[{"name":"Integrated BioTherapeutics, Inc., Rockville, Maryland, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7450-6624","authenticated-orcid":true,"given":"Meagan","family":"McMahon","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4121-776X","authenticated-orcid":false,"given":"Florian","family":"Krammer","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, USA"},{"name":"Center for Vaccine Research and Pandemic Preparedness (C-VaRPP), Icahn School of Medicine at Mount Sinai, New York, New York, USA"},{"name":"Department of Pathology, Molecular and Cell-Based Medicine, Icahn School of Medicine at Mount Sinai, New York, New York, USA"},{"name":"Ignaz Semmelweis Institute, Interuniversity Institute for Infection Research, Medical University of Vienna, Vienna, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4863-0015","authenticated-orcid":false,"given":"Jonathan R.","family":"Lai","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Albert Einstein College of Medicine, Bronx, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.R110.129809"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.3390\/v11050458"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1101\/cshperspect.a028845"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41572-018-0002-y"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01639-18"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2014.04.009"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1008411"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2015.04.028"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1038\/srep45043"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1007233"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.43764"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1080\/21645515.2021.1886560"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1225416"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1111\/imr.12005"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1038\/ni.1746"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2018.12.004"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2017.04.037"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature18938"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1038\/ni.3058"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2021.07.006"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2100104118"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2018.07.009"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1038\/srep19193"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.aag3267"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2021.729851"},{"key":"e_1_3_4_27_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2021.05.001"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.1080\/21645515.2016.1251536"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1222908"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.aay0678"},{"key":"e_1_3_4_31_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.02281-16"},{"key":"e_1_3_4_32_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1304985110"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.1080\/19420862.2016.1148850"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00215-10"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.RA117.001627"},{"key":"e_1_3_4_36_2","doi-asserted-by":"publisher","DOI":"10.1080\/19420862.2016.1270492"},{"key":"e_1_3_4_37_2","doi-asserted-by":"publisher","DOI":"10.1126\/scitranslmed.3009655"},{"key":"e_1_3_4_38_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1220145110"},{"key":"e_1_3_4_39_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-021-03461-y"},{"key":"e_1_3_4_40_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2016.05.024"},{"key":"e_1_3_4_41_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.aan8630"},{"key":"e_1_3_4_42_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0260954"},{"key":"e_1_3_4_43_2","doi-asserted-by":"publisher","DOI":"10.3791\/2057"},{"key":"e_1_3_4_44_2","doi-asserted-by":"publisher","DOI":"10.3791\/51112"},{"key":"e_1_3_4_45_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.03617-20"},{"key":"e_1_3_4_46_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jim.2007.01.008"}],"updated-by":[{"DOI":"10.1128\/mbio.00278-25","type":"correction","label":"Correction","source":"publisher","updated":{"date-parts":[[2025,3,4]],"date-time":"2025-03-04T00:00:00Z","timestamp":1741046400000}}],"container-title":["mBio"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/mbio.01085-24","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/mbio.01085-24","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,7,17]],"date-time":"2024-07-17T09:01:30Z","timestamp":1721206890000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/mbio.01085-24"}},"subtitle":[],"editor":[{"given":"Margaret E.","family":"Ackerman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2024,7,17]]},"references-count":45,"journal-issue":{"issue":"7","published-print":{"date-parts":[[2024,7,17]]}},"alternative-id":["10.1128\/mbio.01085-24"],"URL":"https:\/\/doi.org\/10.1128\/mbio.01085-24","relation":{},"ISSN":["2150-7511"],"issn-type":[{"value":"2150-7511","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,7,17]]},"assertion":[{"value":"2024-04-08","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-05-06","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-06-20","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e01085-24"}}